TGFßR2 EXTRACELLULAR DOMAIN TRUNCATED MOLECULE, FUSION PROTEIN OF TGFßR2 EXTRACELLULAR DOMAIN TRUNCATED MOLECULE AND ANTI-EGFR ANTIBODY, AND ANTI-TUMOR USE OF FUSION PROTEIN

ABSTRACT

Provided are multiple types of TGFβR2 in truncated forms and a fusion protein constructed by TGFβR2 and EGFR antibody HPA8; also provided are a nucleic acid (comprising heavy/light chain variable regions) encoding the antibody, a vector, a pharmaceutical composition, and a kit comprising the nucleic acid; further provided is a fusion protein of the prepared truncated TGFβR2 receptor protein and a targeted EGFR and other multiple types of tumor target antibodies.

CROSS-REFERENCING OF RELATED APPLICATIONS

This application claims the benefit of Chinese patent application 202010351280.6 filed on Apr. 28, 2021, the contents of which are incorporated herein by reference.

FIELD

The present invention relates to the field of tumor immunotherapeutic agents. Specifically, the invention relates to a truncated TGFβR2 (an immunomodulatory factor)extracellular domain molecule, a fusion protein comprising the truncated TGFβR2 extracellular domain molecule, and the targeting portion. The present invention comprises pharmaceutical compositions and their application as antitumor medicaments.

BACKGROUND

Transforming growth factor-β0 (TGF-β) belongs to the TGF-β superfamily that regulates cell growth and differentiation, and is a pleiotropic and multifunctional cytokine that regulates cell proliferation, differentiation, and apoptosis through cell surface receptor signaling pathways in an autocrine or paracrine manner, and plays an important regulatory role in the synthesis of extracellular matrix, repair of trauma, and immune function, etc. Three isoforms, TGF-β1, TGF-β2, and TGF-β3, are present in mammals, and the most abundant and expressed isoform is TGF-β1. TGF-β initiates classical Smad and non-Smad pathways for downstream signaling by binding to TGFβR1 and TGFβR2 serine-threonine kinase receptors on the cell membrane surface.^([1]). In normal homeostasis in vivo, TGF-β signaling regulates key processes such as growth, regeneration, differentiation, etc. of various tissues. In the immune system, TGF-β triggers tolerance and suppresses inflammation. Genetic mutations can alter the TGF-β signaling in tumor-initiating cells. In the initial stages of tumorigenesis, TGF-β plays a key cancer-inhibiting role by inhibiting cell proliferation and triggering the apoptotic program. However, as tumors progress, selective pressure leads to the loss of the tumor suppressive function of TGF-β by tumor cells through different mechanisms. Acquired inactivating mutations in the TGF-β signaling pathway allow a variety of malignant cells to grow in a TGF-β-enriched environment. In addition, tumor cells somehow convert the pro-apoptotic capacity of TGF-β into pro-tumor developmental functions, such as invasion and migration capacity, and promotion of mesenchymal transition.^([2-4]) TGF-β regulates a variety of immune cell types and functions. TGF-β controls adaptive immunity by directly promoting the proliferation of Treg cells thus inhibiting the production and function of effector T cells and antigen-presenting dendritic cells. TGF-β also inhibits NK cell function and converts macrophages and neutrophils into pro-tumor growth subtypes, promoting the formation of a tumor microenvironment with negative tumor immune regulation.^([4]) TGF-β1 is often expressed at higher levels than normal paraneoplastic tissue in a variety of solid tumors including EGFR-positive colorectal cancer, non-small cell lung cancer, and head and neck squamous cell carcinoma. Clinical data suggest that blocking the TGF-β pathway alone is not sufficient to fully restore the immune system to suppress tumorigenesis, and therefore no TGF-β antibodies are yet available.

Epidermal growth factor receptor (EGFR) is an expression product of the proto-oncogene C-ErbB1, a receptor for cell proliferation and signaling of epithelial growth factor (EGF), a member of the epidermal growth factor receptor (HER) family, and a tyrosine kinase receptor with a molecular weight of 170 kDa.^([5]) EGFR is divided into an extracellular ligand-binding region, a transmembrane region, and an intracellular kinase region. The extracellular domain of EGFR is transformed from monomer to dimer after binding to the ligand, activates the intracellular kinase region and multiple downstream signaling pathways, and plays an important role in cell growth, proliferation and differentiation.^([6]) High expression of EGFR causes enhanced downstream signaling, increased expression of mutant EGFR receptors or ligands leads to sustained activation of EGFR, enhanced activity of the secretory loop and disruption of receptor down-regulation mechanism, etc. in turn activates genes related to tumor proliferation and differentiation and plays an important role in tumor formation and development.^([7]) EGFR overexpression is associated with reduced survival in several cancer types, including head and neck, bladder, ovarian, cervical, and esophageal cancers. In addition, anti-EGFR medicaments have been shown to be effective in the treatment of several types of solid tumors such as colorectal, head and neck, non-small cell lung (NSCLC), and pancreatic cancers, including overall survival, progression-free survival, and overall response rates^([8]). Therefore, as a clear target associated with tumor proliferation, EGFR-targeted agents have become the first-line treatment option for a variety of malignancies.

Although the signals initiated by TGF-β are different from those initiated by the EGF/EGFR pathway, the signaling pathways between the two can interact. TGF-β and EGFR signaling have been found to interact and jointly promote tumor progression in a variety of tumors. TGF-β can induce EGFR trans-activation in a highly cell type and context-specific mode. For example, TGF-β promotes the migration and invasion of breast cancer cells (MDA-MB-231, T47D, 4T1) by upregulating EGFR through the classical Smad and ERK/Sp1 signaling pathways^([9, 10]). In squamous cell carcinoma (A431, SCC13), TGF-β activates the EGFR pathway through H₂O₂-dependent mechanisms to increase Erk1/2 phosphorylation levels^([11]). EGF and its associated downstream signaling pathways can also regulate TGF-β signaling in different cell types. For example, in human primary ovarian cancer cells, EGF reduces the sensitivity of ovarian cancer cells to the anti-proliferative effects of TGF-β by decreasing the mRNA expression of the TGF-β-inducible cell cycle regulator p15^(INK4B[12]). Oncogenic Ras in breast and lung epithelial cells reduces the cell growth inhibitory effect of TGF-β by inhibiting TGF-β-mediated signaling via negative regulation of Smad2 and Smad3^([13, 14]) EGF also positively regulates Smad2 signaling in COS7 cells by increasing Smad2 phosphorylation through the ERK pathway^([15]).

TGF-β and EGF synergistically promote the malignant phenotype of tumors, and studies on different tissues have shown that EGF combined with TGF-β enhances epithelial to mesenchymal transition (EMT). For example, EGF and TGF-β1 promote the expression of laminin-332 (Laminin-332) to synergistically promote EMT in oral epithelial cancer.^([16]) EGF and TGF-β1 promote EMT in intestinal epithelial cells by downregulating E-cadherin via the MAPK pathway rather than PI3K, p38MAPK, INK, or AP-1 pathway^([17]). EGF and TGF-β1 induce Slug and Snail expression via Smad and MEK1/2-dependent signaling pathways, downregulate E-cadherin, and promote EMT in ovarian epithelial cells^([18]). EGF and TGF-β1 activate the ERK1/2 signaling pathway, synergistically upregulate Snail protein expression, and promote EMT and migration in human renal cortical proximal tubule epithelial cells (HK-2)^([20]). EGF enhances TGF-β-induced EMT in lung cancer (H322, H358) and pancreatic cancer (HPAF-II, CAPAN-2) cells by promoting the binding of SHP2 to GAB1.^([20]).

Several clinical studies have shown that elevated TGF-β levels are closely associated with the development of medicament resistance and poor prognosis. TGF-β1-induced EMT in osteosarcoma cancer stem-like cells decreases miR-499a expression, leading to increased SHKBP1 expression occurring concomitantly with a TGFβ-induced EMT-associated kinase switch to an AKT-activated EGFR-independent state, thereby reducing EGFR activity and inducing osteosarcoma resistance to EGFR kinase inhibitors (FIG. 5 ).^([21]). Treg cells are one of the main cells that produce TGF-β. The increased number of Treg in tumors of patients with head and neck squamous carcinoma treated with Cetuximab is accompanied by an increase in TGF-β content while the TGF-β content in patients with poor Cetuximab efficacy is even high^([22]). Elevated TGF-β may induce resistance to EGFR antibody therapy by inhibiting the expression of relevant molecular effectors of effector cell-mediated cytotoxicity, activating the EGFR-independent AKT pathway and enhancing EMT inducition.^([23]) In EGFR mutant non-small cell lung cancer, the classical TGF-β/Smad signaling pathway is involved in the development of PD-L1-induced tumor resistance to EGFR kinase inhibitors.^([24]). In breast cancer tissues, TGF-β expression was positively correlated with EGFR expression, and elevated levels of TGF-β and EGFR were associated with poor prognosis in breast cancer patients^([9]). In summary, EGFR and TGF-β play relatively independent and closely related roles in the process of tumor development. In addition, TGF-β is a key molecule in the development of acquired resistance in tumors against EGFR-targeted therapy. Animal studies have shown that inhibition of TGF-β improves the in vivo antitumor effect of Cetuximab on head and neck squamous cell tumor xenografts^([23]). These provide a theoretical basis for combining targeting TGF-β to enhance the therapeutic efficacy of EGFR antibodies against EGFR-positive tumors.

The present invention provides novel fusion proteins comprising truncated TGFβR2 that can specifically target both EGFR and TGF-β, two relatively independent and closely related signaling pathways. It is used to treat solid tumors including but not limited to gastric cancer.

SUMMARY

In one aspect, the present invention provides a truncated TGFβR2 extracellular domain molecule that, compared to its natural form,

a) at least the amino acid residues at positions 6-16 thereof are deleted, and further optionally, the amino acid residues at positions 17-17+n thereof are deleted, where n is an integer from 1-10; preferably, n is 2, 4, 8, 9 or 10; most preferably, n is 9; or

b) on the basis of the deletion of amino acid residues thereof at positions 6-26, furthermore, the amino acid residues thereof at positions 5, 4-5, 3-5, 2-5, 1, 1-2, 1-3, or 1-4 are deleted; or

c) the amino acid residues at positions 7-26 are deleted.

In one embodiment, the amino acid sequence comprises any of SEQ ID NO: 48-62.

In another aspect, the present invention provides a fusion protein comprising the molecules described herein.

In one embodiment, said fusion protein comprises

a) said truncated TGFβR2 extracellular domain molecule and

b) a targeting portion.

In one embodiment, said fusion protein targeting portion is a cancer cell-specific targeting portion selected from an antibody or antigen-binding fragment thereof, a functional ligand or Fc fusion protein thereof, and a receptor protein or Fc fusion protein thereof.

In one embodiment, said fusion protein targeting portion is an anti-EGFR antibody or antigen-binding fragment thereof.

In one embodiment, the N-terminal of the truncated TGFβR2 extracellular domain molecule in a said fusion protein is linked to the C-terminal of the heavy chain of the targeting portion, optionally, by a linker.

In one embodiment, said linker is preferably a G₄ S flexible peptide linker, preferably a (G₄ S)₄ peptide linker.

In a further aspect, the present invention provides an isolated binding antibody or an antigen-binding fragment thereof to EGFR, comprising

(a) a heavy chain variable region comprising heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3 domains comprising SEQ ID NOs: 19, 20, and 21, respectively, and/or

(b) a light chain variable region comprising a light chain CDR1, light chain CDR2, and light chain CDR3 domain comprising SEQ ID NOs: 16, 17, and 18, respectively.

In one embodiment, said antibody or antigen-binding fragment thereof comprises

a) a heavy chain variable region comprising a sequence comprising SEQ ID NO:28 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and/or

(b) a light chain variable region comprising a sequence comprising SEQ ID NO: 29 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith.

In one embodiment, said antibody further comprises:

a) a heavy chain constant region, comprising a sequence comprising SEQ ID NO: 30 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and/or

b) a light chain constant region, comprising a sequence comprising SEQ ID NO: 31 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith.

In one embodiment, the targeting portion of the said fusion protein is selected from said anti-EGFR antibody, Trastuzumab, Bevacizumab, Ramucirumab, Ipilimumab, or Panitumumab.

In one embodiment, said fusion protein comprises

a) the amino acid sequence of a heavy chain comprises SEQ ID NO:141 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and

b) the amino acid sequence of a light chain comprising SEQ ID NO:23 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith.

which comprises two heavy chains and two light chains; a disulfide bond is formed between a first light chain and a first heavy chain thereof, a disulfide bond is formed between a second light chain and a second heavy chain thereof, and a disulfide bond is formed between a first heavy chain and a second heavy chain thereof.

In one embodiment, said fusion protein has a KD value of 2.92 pM-26.3 pM, preferably 7 pM-9 pM, most preferably 8.77 pM, for binding affinity to human EGFR protein; and has a KD value of 23 pM-288.3 pM, preferably 64 pM-144 pM, most preferably 96.1 pM for binding affinity to human TGF-β1 protein.

In a further aspect, the present invention provides a conjugate comprising the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or the antigen-binding fragment thereof as described herein, and an additional therapeutic agent, preferably said antibody or antigen-binding fragment thereof and the additional therapeutic agent are linked by a linker.

In a further aspect, the present invention provides a nucleic acid encoding the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, which is mRNA and/or DNA.

In one embodiment, said nucleic acid comprises

any one of SEQ ID NOs: 32 to 39;

any one of SEQ ID NOs: 67-84; or

any one of SEQ ID NOs: 148-163, or an functional variant thereof.

In a further aspect, the present invention provides an expression vector comprising the nucleic acid described herein.

In a further aspect, the present invention provides a host cell comprising the nucleic acid as described herein or the expression vector as described herein.

In a further aspect, the present invention provides a method for producing the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, comprising culturing the host cell as described herein under conditions suitable for expression of the preceding protein molecule, and recovering the expressed product from the culture medium.

In a further aspect, the present invention provides a pharmaceutical composition comprising

a) the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, or the expression vector as described herein; and

b) a pharmaceutically acceptable carrier; optionally

c) one or more other therapeutic agents.

In a further aspect, the present invention provides use of the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, the expression vector as described herein, or the pharmaceutical composition as described herein for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.

In a further aspect, the present invention provides use of the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, the expression vector as described herein, or the pharmaceutical composition as described herein for the preparation of a medicament for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.

In a further aspect, the present invention provides a pharmaceutical combination comprising the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, the expression vector as described herein, or the pharmaceutical composition as described herein, and one or more additional therapeutic agent(s).

In a further aspect, the present invention provides a kit comprising the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, the expression vector as described herein, or the pharmaceutical composition as described herein; preferably, further comprising a device for administering a medicament.

In a further aspect, the present invention provides a method of preventing and treating a neoplastic disease comprising administering to a subject the molecule of the truncated TGFβR2 extracellular domain molecule as described herein, the fusion protein as described herein, the antibody or an antigen-binding fragment thereof as described herein, the conjugate as described herein, the nucleic acid as described herein, the expression vector as described herein, or the pharmaceutical composition as described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 Binding assay of murine antibody EGFR-mhPA8 to recombinant human EGFR-His protein.

FIG. 2 Blockage of Blockage of EGF binding to EGFR by the murine antibody EGFR-mhPA8.

FIG. 3 Inhibition of proliferation of MDA-MB-468 cells by murine antibody EGFR-mhPA8.

FIG. 4 Binding assay of humanized antibody EGFR-HPA8 to recombinant human EGFR-His protein.

FIG. 5 Blockage of Blockage of EGF binding to EGFR by humanized antibody EGFR-HPA8.

FIG. 6 Inhibition of proliferation of MDA-MB-468 cells by humanized antibody EGFR-HPA8 under different ligand conditions.

FIG. 7 Inhibition of proliferation of Fadu cells by humanized antibody EGFR-HPA8 under different ligand conditions.

FIG. 8 Effect of ADCC mediated by humanized antibody EGFR-HPA8.

FIG. 9 Effect of EGFR-HPA8 on the body weight of gastric cancer cell SNU-5 subcutaneously xenografted mice.

FIG. 10 Effect of EGFR-HPA8 on tumor volume and TGI results of gastric cancer cell SNU-5 subcutaneously xenografted mice.

FIG. 11 Effect of EGFR-HPA8 on the body weight of non-small cell lung cancer NCI-H1975 cell subcutaneously xenografted mice.

FIG. 12 Effect of EGFR-HPA8 on tumor volume and TGI results in non-small cell lung cancer NCI-H1975 cell subcutaneously xenografted mice.

FIG. 13 Schematic structure of EGFR antibody/TGFβR2 fusion protein.

FIG. 14 Detection of degradation of EGFR antibody/TGFβR2 fusion protein.

FIG. 15 Detection of degradation propensity of EGFR antibody/TGFβR2 fusion protein.

FIG. 16 Detection of the binding ability of different truncated forms of EGFR antibody/TGFβR2 fusion protein to TGF-β1 and EGFR.

FIG. 17 Detection of the ability of EGFR antibody/TGFβR2 fusion protein TGFβR2 truncated EGFR antibody/TGFβR2 fusion protein to neutralize TGF-β1.

FIG. 18 Detection of the ability of EGFR antibody/TGFβR2 fusion protein TGFβR2 truncated EGFR antibody/TGFβR2 fusion protein to neutralize TGF-3.

FIG. 19 Assay of the ability of fusion protein 6 to bind to TGF-β1 and TGF-β3 proteins respectively.

FIG. 20 Assay of the ability of fusion protein 6 to block TGF-β1 protein or TGF-β3 protein from binding TGFβR2-Fc respectively.

FIG. 21 Assay of the binding and competition ability of fusion protein 6.

FIG. 22 Affinity assay of fusion protein 6 for recombinant human EGFR protein.

FIG. 23 Affinity assay of fusion protein 6 for recombinant human TGF-β1 protein.

FIG. 24 Neutralization effects of TGF-β1 by fusion protein 6 on Mv-1-lu cells.

FIG. 25 Neutralization effects of TGF-β1 by fusion protein 6 assayed by the reporter gene system.

FIG. 26 Inhibition of proliferation of MDA-MB-468 cells by fusion protein 6.

FIG. 27 Fusion protein 6-mediated effects of ADCC.

FIG. 28 Effect of fusion protein 6 on the body weight of non-small cell lung cancer NCI-H1975 cell subcutaneously xenografted mice.

FIG. 29 Effect of fusion protein 6 on tumor volume in non-small cell lung cancer NCI-H1975 cell subcutaneously xenografted mice.

FIG. 30 Ultrafiltration stability assay of fusion protein 6.

FIG. 31 Assay of degradation of X/TGFβR2 fusion protein.

FIG. 32 Assay of degradation propensity of X/TGFβR2 fusion protein.

FIG. 33 Assay of the binding capacity of X/TGFβR2 fusion protein to TGF-β1.

FIG. 34 Assay of the ability of X/TGFβR2 fusion protein to neutralize TGF-β1 and TGF-β3.

FIG. 35 Assay of X/TGFβR2 fusion protein binding to X-portion target.

DETAILED DESCRIPTION Definition

Unless otherwise stated, all technical and scientific terms used herein have the meaning commonly understood by those of skilled persons in the art to which the present invention belongs. For the purposes of the present invention, the following terms are further defined.

When used herein and in the appended claims, the singular forms “one”, “a”, “another”, and “said” include the plural designation of the object unless the context clearly indicates otherwise.

The term “truncated” form of a protein molecule means in which one or more amino acid residues are missing in a modified manner compared to the natural form of the protein molecule.

The term “fusion protein” refers to a protein molecule that combines two or more proteins. It is usually obtained by expressing a hybrid gene that combines the sequences of two or more genes, which are inserted into the expression vector in a form that conforms to the expression frame.

The term “antibody” refers to an immunoglobulin molecule and refers to any form of antibody that exhibits the desired biological activity. It includes, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies and multi-specific antibodies (e.g., bispecific antibodies), and even antibody fragments.

The term “variable region” refers to the domain in the heavy or light chain of an antibody that is involved in antibody binding to antigen. The variable regions of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have a similar structure and can be further subdivided into highly variable regions (called complementary decision regions (CDR)) interspersed with more conserved regions (called framework regions (FR)).

The complementarity determining region (CDR) and the framework region (FR) of a given antibody can be identified using the Kabat system (Kabat et al: Sequences of Proteins of Immunological Interest, 5th edition, U.S. Department of Health and Human Services, PHS, NIH, NIH Publication No. 91-3242, 1991).

The term “constant region” refers to such amino acid sequences in the light and heavy chains of an antibody that is not directly involved in antibody-antigen binding but exhibit a variety of effector functions, such as antibody-dependent cytotoxicity.

An “antigen-binding fragment of an antibody” comprises a portion of an intact antibody molecule, retains at least some of the binding specificity of the parent antibody, and typically includes at least a portion of the antigen-binding region or variable region (e.g., one or more CDRs) of the parent antibody. Examples of antigen-binding fragments include, but are not limited to, Fv, Fab, Fab′, Fab′-SH, F(ab′)2, Fd fragments, Fd′ fragments, single-chain antibody molecules (e.g., scFv, di-scFv or tri-scFv, bipartite antibodies or scFab), and single-domain antibodies.

The term “conjugate” refers to a biologically active protein or peptide molecule that forms a covalent or non-covalent linkage with another molecule, either a small molecule or a biomolecule.

The term “monoclonal antibody” refers to an antibody derived from a substantially homogeneous population of antibodies, i.e., said population comprising homogeneous antibodies identical to each other except for possible mutations (e.g., natural mutations) that may be present in very small amounts. Thus, the term “monoclonal” indicates the nature of said antibodies, i.e., not a mixture of unrelated antibodies. In contrast to polyclonal antibody preparations, which usually include different antibodies against different decision clusters (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a separate decision cluster on the antigen. In addition to their specificity, monoclonal antibody preparations have the advantage that they are usually not contaminated with other antibodies. The term “monoclonal” should not be construed as requiring any particular method of producing said antibody. The term monoclonal antibody specifically includes chimeric antibodies, humanized antibodies, and human antibodies.

The antibody “specifically binds” to a target antigen such as a tumor-associated antigen protein (herein, EGFR), i.e., binding said antigen with sufficient affinity to allow said antibody to be used as a therapeutic agent, targeting a tissue or cell expressing said antigen, and without significantly cross-reactivity with other proteins or with proteins other than the homologs and variants (e.g., mutant forms, splice variants, or protein hydrolysis truncated forms) of the antigen targets mentioned above.

The term “binding affinity” refers to the strength of the sum of non-covalent interactions between a molecule's individual binding sites and its binding partners. Unless otherwise specified, “binding affinity” as used herein refers to the intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). “K_(D)”, “binding rate constant kon” and “dissociation rate constant koff” are commonly used to describe the affinity between a molecule (e.g., antibody) and its binding partner (e.g., antigen) affinity, i.e., how tightly a ligand binds a particular protein. The binding affinity is influenced by non-covalent intermolecular interactions such as hydrogen bonding, electrostatic interactions, and hydrophobic and van der Waals forces between two molecules. In addition, the binding affinity between a ligand and its target molecule may be influenced by the presence of other molecules. Affinity can be analyzed by conventional methods known in the art, including the ELISA described herein.

The term “targeting portion” refers to the portion of a fusion protein that has the function of specifically binding to a target cell. The term includes antibodies and other natural (e.g., receptors, ligands) or synthetic (e.g., DARPin) molecules that specifically bind to target cells. As used herein, “specifically binds target cells” indicates that said portion preferentially binds target cells within a complex mixture.

An “isolated” biomolecule is a biomolecule that has been identified and isolated from a cell that naturally expresses the molecule. Isolated biomolecules include in situ biomolecules in recombinant cells as well as biomolecules that are typically prepared by at least one purification step.

The term “receptor” is a biochemical concept that refers to a class of molecules that specifically recognize and bind extracellular signals (i.e., the term “ligand”) and produce a specific effect within the cell. The effects produced may last only for a short period of time, such as altering cell metabolism or cell motility. It may also be a long-lasting effect, such as up- or down-regulating the expression of a gene or genes.

“Antibody-dependent cell-mediated cytotoxicity” or “ADCC” refers to a form of cytotoxicity wherein secreted Ig binding to Fcγ receptors present on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages) enables these cytotoxic effector cells to specifically bind to antigen-bearing target cells and subsequently kill said target cells using, for example, a cytotoxic agent. To assess the ADCC activity of the target antibody, an in vitro ADCC assay can be performed, such as the in vitro ADCC assay documented in U.S. Pat. No. 5,500,362 or 5,821,337 or U.S. Pat. No. 6,737,056 (Presta), the methods documented in embodiments of this application. Useful effector cells for such assays include PBMCs and NK cells.

A truncated TGFβR2 extracellular domain molecule of the present invention, fusion proteins comprising the truncated TGFβR2 extracellular domain molecule, and the targeting portion targeting portion

Multiple susceptibility sites exist between the N-terminal sites 7-15 of the full-length TGFβR2 extracellular domain. In order to improve the structural stability of the fusion protein, the inventors modified the N-terminal amino acid sequence of the extracellular domain of TGFβR2 by deletion of amino acids in deferent numbers. Fifteen truncated TGFβR2 extracellular domain molecules were obtained.

The inventors used their newly isolated EGFR antibodies EGFR-HPA8, Trastuzumab, Bevacizumab, Ramucirumab, Ipilimumab, or Panitumumab as the targeting portion of the fusion protein and the truncated TGFβR2 protein extracellular domain molecule as the immunomodulatory portion of the fusion protein. The truncated TGFβR2 protein extracellular domain molecule is linked to the C-terminal of the heavy chain of EGFR antibody by homologous recombination method, the fusion protein (EGFR/TGFβR2) comprising two chains, i.e. the light chain and heavy chain of the EGFR antibody and TGFβR2 extracellular domain, is formed, the structure of which is shown in FIG. 13 . In EGFR antibody/TGFβR2 fusion protein, the amino acid of the heavy chain C-terminal of the EGFR antibody links to the extracellular domain of TGFβR2 of different amino acid deletion forms by (G₄S) 4 Linker (SEQ ID NO:66). In addition, the lysine residue of the heavy chain C-terminal of the EGFR antibody is removed to reduce the risk of proteolysis.

In a preferred embodiment of the present invention, fusion protein 6 retains a high binding affinity for human EGFR protein, having a K_(D) value of 8.77 pM for binding affinity for human EGFR protein, a binding constant kon value of 1.68E+06 M⁻¹ s⁻¹, and a dissociation constant kdis of 1.47E-05 s⁻¹. In addition, fusion protein 6 which has the truncated TGFβR2,has a similar affinity for human TGF-β1 protein as fusion protein 1, which has full-length TGFβR2, with a k_(D) value of 96.1 pM, a binding constant kon value of 1.53E+06 M⁻¹ s⁻¹, and a dissociation constant kdis of 1.47E-04 s⁻¹.

Nucleic Acids of the Present Invention

The invention also relates to nucleic acids encoding the truncated TGFβR2 extracellular domain molecule of the present invention, molecules including fusion proteins comprising the truncated molecule and the targeting portion targeting portion, or portions thereof. Some example sequences of these nucleic acid molecules are shown in the sequential list.

The nucleic acid molecules of the present invention are not limited to the sequences disclosed herein, but also include variants and other corresponding nucleic acid forms thereof, such as mRNA, cDNA, and variants thereof. The variants of the present invention may be described with reference to their physical properties in hybridization. Those of skill in the art will recognize that using nucleic acid hybridization techniques, nucleic acids can be used to identify their complements as well as their equivalents or congeners.

Recombinant Vectors and Expression

The present invention also provides recombinant constructs comprising one or more nucleotide sequence(s) of the present invention. The recombinant constructs of the present invention may be used with vectors, for example, plasmid, phagemid, phage, or viral vectors, and nucleic acid molecules encoding antibodies of the present invention are inserted into said vectors.

The present truncated TGFβR2 extracellular domain molecule, targeting portion targeting portions, fusion proteins comprising the truncated molecule and the targeting portion targeting portion can be prepared by the recombinant expression of a nucleotide sequence encoding said molecule or protein in a host cell. Said molecule or protein may contain more than one amino acid sequence, and in order to express the plurality of amino acid sequences by recombinant methods, one or more recombinant expression vectors carrying the encoding nucleotide sequence may be transfected with the host cell so that said plurality of amino acid sequences is expressed in the said host cell. Standard recombinant DNA methodologies are used to prepare and/or obtain nucleic acids encoding said molecules or proteins, incorporate these nucleic acids into recombinant expression vectors and introduce said vectors into host cells, e.g. Sambrook, Fritsch, and Maniatis (eds.), Molecular Cloning; A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y., (1989), Ausubel, F. M. et al. (eds.) Current Protocols in Molecular Biology, Greene Publishing Associates, (1989) and those documented in U.S. Pat. No. 4,816,397 by Boss et al.

Examples of prokaryotic host cells are bacteria and examples of eukaryotic host cells are yeast, insect, or mammalian cells. It should be understood that the design of an expression vector including the selection of regulatory sequences is influenced by a variety of factors, such as the selection of the host cell, the level of expression of the desired protein, and whether the expression is constitutive or inducible.

The truncated TGFβR2 extracellular domain molecules, targeting portion targeting portion and fusion proteins of the present invention can be recovered and purified from recombinant cell cultures by well-known methods, said well-known methods including, but not limited to, ammonium sulfate or ethanol precipitation, acid extraction, protein A affinity chromatography, protein G affinity chromatography, anion or cation exchange chromatography, cellulose phosphate chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxyapatite chromatography, and lectin chromatography. High-performance liquid chromatography (“HPLC”) can also be used for purification.

Use

The truncated TGFβR2 extracellular domain molecules, antibodies and fusion proteins of the present invention can be used to treat cancers, such as gastric cancer.

Pharmaceutical Compositions

One or more kinds of the truncated molecule(s), fusion proteins, antibodies, and antigen-binding fragments, truncated molecule-medicament conjugates, fusion protein-medicament conjugates, conjugates, nucleic acids, and carriers of the present invention may be prepared with at least one other chemical agent to form a pharmaceutical composition comprising the preceding active ingredient(s) described above and one or more medicament-acceptable carriers, diluents, or excipients; optionally, also may also comprise one or more other therapeutic agents.

Reagent Kits

The present invention also relates to pharmaceutical packaging and kits comprising one or more containers, said containers containing the pharmaceutical compositions of the present invention as mentioned above. Associated with such containers may be tipped in the form prescribed by the government agency governing the manufacture, use, or distribution of the medicament or biological product, which reflects approval for human administration by the agency in which said product is manufactured, used, or distributed.

Preparation and Storage

The pharmaceutical compositions of the present invention can be prepared in a manner known in the art, for example by conventional methods of mixing, dissolving, granulating, grinding, emulsifying, encapsulating, encapsulating, or lyophilizing.

After pharmaceutical compositions comprising compounds of the present invention formulated in an acceptable carrier have been prepared, they can be placed in appropriate containers and labeled for the treatment of the indicated condition. Such labeling would include the amount, frequency, and method of administration of the medicament.

Medicament Combinations

The combinations comprising the antibodies of the present invention described above are also combined with one or more other therapeutic agents (s), wherein the resulting combination does not cause unacceptable adverse effects.

The following embodiments are used to illustrate the invention exemplarily and are not intended to limit the invention.

EXAMPLE Example 1: Screening for Murine Antibodies that Block the Binding of EGF to EGFR Using Phage Antibody Display Library

1.1 Mouse Immunization and Phage Antibody Library Screening

Mice were immunized with the nucleic acid plasmids pCMV3-mFlt3L (G12FE5S01-D, constructed by Sinocelltech Limited, SEQ ID NO:142) and pCMV3-mCSF2 (G12FE5S02-D, constructed by Sinocelltech Limited, SEQ ID NO:143). The specific method is as follows: mice were immunized with cardiotoxin 10 M intramuscularly two days before the first immunization. 20 g/leg of pCMV3-mFlt3L and pCMV3-mCSF2 mixture with a mass ratio of 1:1 was injected intramuscularly and 30 g/leg of pGS6-EGFR-TT-WPRE (Constructed by Sinocelltech Limited, SEQ ID NO:144) was injected intramuscularly three days later for each immunization, intraperitoneal injection of 5×10⁶ insect cells overexpressing EGFR for fourth and fifth immunization, with immunization intervals of 2 weeks, 2 weeks, 2 weeks, and 2 weeks, in sequence. From the third immunization onwards, blood was collected via the medial canthal plexus of the eye seven days after each immunization. The recombinant human EGFR protein (source: Sino Biological, Inc., Cat. 10001-H08B, hereinafter the same) was coated to detect the serum titre of anti-EGFR in mice. The fifth immzation serum achieved a criteria that immune serum titers reaching 8000-fold and serum titre OD>1, then an intraperitoneal booster injection using 5×10⁶ MDA-MB468 cells was performed at an interval of 20 days, and 7 days later, the mice were executed and the spleen tissues were frozen in liquid nitrogen.

Mouse spleen tissue was extracted with TriPure Isolation Reagent (source: Roche Cat. No. 11 667 165 001) for RNA extraction, and cDNA was obtained by reverse transcription with the reverse transcription kit TriPure Isolation Reagent (source: Invitrogen Cat. No. 18080-051), PCR amplification was performed to obtain the nucleotide sequences encoding the light and heavy chain variable regions of mouse antibodies, the nucleotide sequences encoding the light and heavy chain variable regions of mouse antibodies were spliced into the nucleotide sequences encoding scFv by overlap extension splicing PCR, light and heavy chain variable regions were linked by linker^([25]):

(SEQ ID NO: 2) TCTAGTGGTGGCGGTGGTTCGGGCGGTG GTGGAGGTGGTAGTTCTAGATCTTCC,

then were ligated into the phage vector pComb3x (source: Sino Biological, Inc.) by restriction endonuclease Sfi I (source: Fermentas), and the phage display scFv antibody library for immunized mice was constructed by electrotransforming X-Blue competent cell. The recombinant human EGFR protein was coated on ELISA plates, and an anti-EGFR positive antibodies enriched phage library were obtained by phage antibody panning process. Subsequently, the above library was mixed with MDA-MB-468 cells (source: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences), and phage libraries enriched for positive antibodies against MDA-MB-468 cells were screened from the above libraries according to the phage antibody panning process (O'Brien, P. M., & Aitken, R. (Eds.), Springer Science & Business Media. 2002; ISBN: 9780896037113). Colonies of phage were selected from the enriched library, expressed, and the binding to recombinant human EGFR protein was detected by ELISA, and a high binding antibody EGFR-mhPA8 scFv antibody clone with specific binding to recombinant human EGFR was screened and was sent to a sequencing company for sequencing to obtain the nucleotide sequence of EGFR-mhPA8 scFv antibody (SEQ ID NO:3).

1.2 Functional Assay of Murine Antibodies Targeting EGFR

1.2.1 Murine Antibody Binding and Ligand Competition Functions

ELISA was used to detect the binding ability of the murine antibody to recombinant human EGFR protein. Recombinant human EGFR-His protein (source: Sino Biological, Inc., hereafter the same) at different concentrations (1.37 ng/mL, 4.12 ng/mL, 12.35 ng/mL, 37.04 ng/mL, 111.11 ng/mL, 333.33 ng/mL, 1000 ng/mL and 3000 ng/mL) were coated in 96-well plates, 100 L per well, were coated overnight at 4° C. The plates were washed the next day and blocked at room temperature for 1 h. EGFR-mhPA8 and the negative control antibody H7N9-R1 (source: Sinocelltech Limited, same below) were added at 13.89 nM and incubated for 1 h, after which the plates were washed to remove the unbound antibody. The secondary antibody Goat anti-hIgG F(ab)2/HRP (source: Jackson ImmunoResearch, Cat. 109-036-006, same as below) was added and incubated for 1 h, the plates were repeatedly washed and the substrate chromogenic solution was added for color development, and the OD₄₅₀ is read by a microplate reader after termination. Using the recombinant human EGFR-His protein concentration as the horizontal coordinate and the OD₄₅₀ reading as the vertical coordinate, the S-curve was fitted and the EC₅₀ of antibody binding to recombinant human EGFR-His protein was analyzed using GraphPad Prism 6.0 software. The results are shown in FIG. 1 . The human-mouse chimeric antibody EGFR-mhPA8 can effectively bind recombinant human EGFR-His with an EC₅₀ of 128.7 ng/mL and R²=1.000. Negative control H7N9-R1 did not bind to recombinant human EGFR-His protein.

This example further analyzed the ability of EGFR-mhPA8 to block EGFR ligand-receptor binding by FACS. 3×10⁵ MDA-MB-468 cells were added with 10 μL of the final concentration of 217.1 nM biotin-labeled EGF-Fc protein (source: Sino Biological, Inc.). EGFR-mhPA8 antibodies at final concentrations of 306.74 nM, 102.25 nM, 34.08 nM, and 11.36 nM were added after 30 min incubation at 2-8° C., and H7N9-R1 was used as the negative control antibody. Mix and incubate at 2-8° C. for 20 min and then wash with PBS and centrifuge to remove unbound antibodies and ligands. Add Streptavidin-488-FITC secondary antibody (source: Sino Biological, Inc., same below) and incubate for 1 h at 2-8° C. Repeat washing and centrifugation to remove unconjugated secondary antibodies. Finally, 200 μL PBS was added to resuspend the cells and filtered through a 400 mesh filter into a flow tube for flow-through detection. The results are shown in FIG. 2 , EGFR-mhPA8 can effectively block the binding of EGF-Fc protein to EGFR on MDA-MB-468 cells in a concentration gradient.

1.2.2 Inhibition of Proliferation of MDA-MB-468 Cells by Murine Antibodies

Breast cancer MDA-MB-468 cells highly express EGFR and also autocrine various ligand factors of EGFR. In this example, the growth inhibitory function of murine antibody on MDA-MB-468 cells was determined by WST-8 assay.

MDA-MB-468 cells were uniformly inoculated in 96-well plates at 5×10³/well. Cells were incubated in CO₂ incubator for 3 h, and different concentrations of murine antibody EGFR-mhPA8 (66.7 nM, 22.2 nM, 7.4 nM, 2.5 nM, 0.82 nM, 0.27 nM, 0.093 nM, 0.033 nM and 0.013 nM) were added. A negative control M group (containing cells) and a blank control B group (medium only, no cells) were also used. Cells were incubated in CO₂ incubator at 37° C. and 5% CO₂ for 5 days and then WST-8 was added at 15 μL/well. After 240 min of incubation, OD₄₅₀-OD₆₃₀ was measured by a microplate reader, and the inhibition rate of mouse-derived antibody was calculated by subtracting the reading value of detection blank well B. The inhibition rate=(OD of group M−OD of the sample)/(OD of group M)×100%, and the quantitative efficacy curve was analyzed and plotted using GraphPad Prism software, the horizontal coordinate is the antibody concentration and the vertical coordinate is the inhibition rate. As shown in FIG. 3 , EGFR-mhPA8 effectively inhibited the proliferation of MDA-MB-468 and the inhibition rate increased with increasing medicament concentration in an “S” curve. EGFR-mhPA8 inhibited MDA-MB-468 cells with EC₅₀ at 2.78 nM and R²=0.991.

Example 2: Humanization of Murine Antibody EGFR-mhPA8 and Ex Vivo Characterization of Humanized Antibody

2.1 Humanization and Production of Murine Antibody mhPA8

The nucleotide sequence of the EGFR-mhPA8 antibody was deduced to obtain the heavy chain (SEQ ID NO:8) and light chain variable region amino acid sequence (SEQ ID NO:9) of the EGFR-mhPA8 scFv antibody. The amino acid sequences of three CDRs each for the EGFR-mhPA8 scFv light and heavy chains were determined by reference to Kabat [26] and the IMGT numbering approach, see SEQ ID NO:10-15. According to Kabat numbering, except for the N mutation to D at position 52 in LCDR2, respective three CDRs of the above-mentioned light and heavy chains were transplanted in the subsequent humanization step and retained in the final humanized antibody EGFR-HIPA8 scFv.

TABLE 1 EGFR-mhPA8 light chain and heavy chain CDR sequences Name Sequence SEQ ID LCDR1 RASENIYYSLA SEQ ID NO: 10 LCDR2 ITNGLAD SEQ ID NO: 11 LCDR3 KQSYDVPLT SEQ ID NO: 12 HCDR1 GYTFTTYYTH SEQ ID NO: 13 HCDR2 WIYPGDVNTKYNEKFKG SEQ ID NO: 14 HCDR3 AREDPGSNYFDY SEQ ID NO: 15

Humanization of murine antibodies was performed using the classical CDR graft method^([27, 28]). The antibodies being of more than 50% amino acid sequence similarity to both the variable regions of light chain and heavy chain of murine antibody, and more than 50% amino acid sequence identity to the framework regions of the variable regions of the light chain and heavy chain of the murine antibodies to be modified respectively were selected as the humanization template library. The human antibody with the highest spatial similarity to the variable region of the antibody to be modified was selected as the humanization template. The three CDR sequences of the light or heavy chains of the murine antibody were substituted with the corresponding CDR amino acid sequences in the humanization template, and the amino acids NG, NS, NA, and NT, which are at high risk of deamidation in the mutated sequences, were subjected to mutation in order to improve the chemical stability and maintain the biological function of the antibody. The affinity of the humanized antibodies is assayed by ELISA and the humanized antibodies that maintain affinity are selected. The humanization template used for the light chain variable region grafted of EGFR-mhPA8 in this example is IGKV1-NL1*01, which has 68.4% homology with the light chain of EGFR-mhPA8, and the humanization template for the heavy chain variable region is IGHV1-69-2*01, which has 64.9% homology with the heavy chain of EGFR-mhPA8. The N52 of humanized antibody EGFR-HPA8 variable region LCDR2 prone to deamidation is mutated to D.

Since the key site of the murine framework region is essential for maintaining the stability of the CDR spatial structure, the key site should be reverse mutated to the corresponding amino acid of the murine antibody. According to the Kabat numbering, the light chain was reverse mutated to Q at position 45, I at position 48, K at position 74, and D at position 76, and the heavy chain was reverted mutated to K at position 38, I at position 48 and L at position 70. The humanized antibody EGFR-HPA8 was obtained by CDR humanization grafting and frame region reversion mutation, and its heavy chain and light chain amino acid sequences are shown in SEQ ID NO:22/23, respectively; its heavy chain and light chain amino acid sequences containing the signal peptide are shown in SEQ ID NO:24/25, respectively, containing the amino acid sequences of sequentially linked heavy chain/light chain signal peptide (SEQ ID NO:26/27), the variable region of the heavy chain/light chain of its humanized antibody (SEQ ID NO:28/29) and the constant region of its humanized antibody, that is, IgG1 heavy chain constant region/human kappa light chain constant region (SEQ ID NO:30/31); and the sequence of its humanized CDR, as detailed in Table 2.

TABLE 2 EGFR-HPA8 light chain and heavy chain CDR sequences Name Sequence SEQ ID LCDR1 RASENIYYSLA SEQ ID NO: 16 LCDR2 ITDGLAD SEQ ID NO: 17 LCDR3 KQSYDVPLT SEQ ID NO: 18 HCDR1 GYTFTTYYTH SEQ ID NO: 19 HCDR2 WIYPGDVNTK SEQ ID NO: 20 YNEKFKG HCDR3 AREDPGSNYFDY SEQ ID NO: 21

The signal peptide containing EGFR-HPA8 antibody light chain nucleotide sequence (SEQ ID NO: 33) containing sequentially linked light chain signal peptide nucleotide sequences (SEQ ID NO: 35), humanized antibody light chain variable region nucleotide sequences (SEQ ID NO: 37) and human kappa light chain constant region nucleotide sequences (SEQ ID NO: 39) was amplified by splicing PCR. The above PCR products were inserted into pSTEP2 vector (source: constructed by Sinocelltech Limited, same below) by In-fusion method (double digested by Hind III+Xba I), and the correct plasmids were verified by sequencing. The nucleotide sequence of the heavy chain variable region of EGFR-HPA8 antibody (SEQ ID NO:36) was obtained by whole gene synthesis and inserted into the pSTEP2 vector (after Sca I+Nhe I double digestion)containing the nucleotide sequence of the heavy chain signal peptide (SEQ ID NO:34) and the nucleotide sequence of the heavy chain constant region of human IgG1 (SEQ ID NO:38) by In-fusion method, and the correct EGFR-HPA8 light-heavy chain expression vector was verified by sequencing. After plasmid extraction and transfection of HEK-293 cells (fut8 knockout), the cells were cultured and expressed for 7 days, the cell supernatant was purified using affinity chromatography after centrifugation, and the chromatography medium was Protein A packing that interacts with Fc. After equilibrating the Protein A chromatography column with 50 mM Tris, 10 mM NaCl, and pH 8.0 buffer for 5-10 times the column volume, the filtered culture supernatant was added to the chromatography column for binding, and the column was drenched with 20 mM Tris, 0.3 M Arg, pH 6.5 buffer for 5-10 times the column volume, and then the column was rinsed with 0.1 M Gly, 10 mM NaCl, pH 3.5 elution buffer, and the collected samples were neutralized with 2 M Tris (pH 8.0) to obtain high-purity and high-quality ADCC-enhanced EGFR-HPA8 antibody.

Primer sequences for splicing PCR amplification of EGFR-HPA8 antibody light chain containing the signal peptide

F1 (SEQ ID NO. 167) GTCACCGTCCTGACAC GAAGCTTGCCGCCACC R1 (SEQ ID NO. 168) ACTATAGAATAGGGCC CTCTAGA F2 (SEQ ID NO. 169) GGCAAGGCTCCAAAGC TGCTGATTTAC R2 (SEQ ID NO. 170) GTAAATCAGCAGCTTT GGAGCCTTGCC F3 (SEQ ID NO. 171) ACCTACTACTGTATGC AGTCCTATGAT R3 (SEQ ID NO. 172) ATCATAGGACTGCATA CAGTAGTAGGT

Primer sequences for whole gene synthesis of EGFR-HPA8 antibody heavy chain variable region

F4 (SEQ ID NO. 173) GCTACCAGGGTGCTGAGTGAGGT CCAACTTGTCCAGTCTGGAGCAG AGGTG R4 (SEQ ID NO. 174) CACTGTGGCTCCAGGCTTCTTCA CCTCTGCTCC F5 (SEQ ID NO. 175) CCTGGAGCCACAGTGAAGATTTC CTGTAAGGTG R5 (SEQ ID NO. 176) GGTGGTGAAGGTGTAGCCAGACA CCTTACAGGA F6 (SEQ ID NO. 177) TACACCTTCACCACCTACTACAC CCACTGGG R6 (SEQ ID NO. 178) CTTGCCAGGAGCCTGCTTCACCC AGTGGGTGT F7 (SEQ ID NO. 179) CAGGCTCCTGGCAAGGGATTGGA GTGGATTG R7 (SEQ ID NO. 180) ATCTCCAGGGTAAATCCAGCCAA TCCACTCCA F8 (SEQ ID NO. 181) ATTTACCCTGGAGATGTGAACAC CAAATACA R8 (SEQ ID NO. 182) CCTGCCCTTGAACTTCTCATTGT ATTTGGTGT F9 (SEQ ID NO. 183) AAGTTCAAGGGCAGGGTGACCCT GACAGCAGAC R9 (SEQ ID NO. 184) ATAGGCTGTGTCTGTGCTGGTGT CTGCTGTCAG F10 (SEQ ID NO. 185) ACAGACACAGCCTATATGGAACT GTCCTCCCTG R10 (SEQ ID NO. 186) GACTGCTGTGTCCTCAGACCTCA GGGAGGACAG F11 (SEQID NO. 187) GAGGACACAGCAGTCTACTACTG TGCCAGGGAG R11 (SEQ ID NO. 188) AAAGTAGTTGCTGCCAGGGTCCT CCCTGGCACA F12 (SEQID NO. 189) GGCAGCAACTACTTTGACTACTG GGGACAAGGC R12 (SEQ ID NO. 190) TGGGCCCTTGGTGCTTGCGCTGG ACACTGTCACCAGGGTGCCTTGT CCCCA

2.2 In Vitro Characterization of Humanized EGFR-HPA8 Antibody

2.2.1 Humanized Antibody-Specific Binding and Ligand Competition Assay

Referring to Example 1.2.1, the binding ability of the human antibody to recombinant human EGFR protein was detected by applying ELISA while setting SCT200 (documented in CN200610012002.8, same below), Erbitux (MERCK, 201621, same below) and negative control. As shown in FIG. 4 , the EC₅₀ for humanized EGFR-HPA8 antibody specifically binding to recombinant human EGFR-His is at 116.6 ng/mL, R²=1.000; EC₅₀ for SCT200 at 166.5 ng/mL, R²=1.000; EC₅₀ for Erbitux at 253.6 ng/mL, R²=1.000; negative control H7N9-R1 did not bind. The results indicated that EGFR-HPA8 had a better ability to bind to recombinant human EGFR-his than SCT200 and Erbitux.

Meanwhile, the ability of humanized antibody EGFR-HPA8 to block EGFR ligand-receptor binding was analyzed by FACS with reference to Example 1.2.1, setting SCT200, Erbitux, and negative control. The results are shown in FIG. 5 , the ability of EGFR-HPA8 to block the binding of EGF-Fc protein to EGFR on MDA-MB-468 cells is stronger than SCT200 and Erbitux.

2.2.2 Inhibition of Different Tumor Cells Proliferation by Humanized Antibodies

2.2.2.1 Ability of Humanized Antibodies to Inhibit the Proliferation of MDA-MB-468 Cells

MDA-MB-468 cells were uniformly inoculated in 96-well plates at 5×10³/well. Cells were incubated in CO₂ incubator for 3 h, and different concentrations of humanized antibody EGFR-HPA8 (666.7 nM, 222.2 nM, 74.1 nM, 24.7 nM, 8.23 nM, 2.74 nM, 0.91 nM, 0.31 nM and 0.1 nM) were added, and SCT200 and Erbitux were used as control. The ligands, HB-EGF (source: Sino Biological, Inc., same below) at a final concentration of 8 ng/mL, BTC-Fc (source: Sino Biological, Inc., same below) at a final concentration of 200 ng/mL or Fc-EREG (source: Sino Biological, Inc., same below) at a final concentration of 1 μg/mL, were then added respectively. Cells were incubated in CO₂ incubator at 37° C. and 5% CO₂ for 5 days, then WST-8 was added at 15 μL/well. After 240 mins of incubation, OD₄₅₀-OD₆₃₀ was measured by a microplate reader, and the growth inhibition rate of cells by the antibody was calculated. The addition of ligand without antibody was used as group M. The inhibition rate=(OD of group M−OD of sample)/(OD of group M)×100%, and the quantitative efficacy curve was analyzed and plotted using GraphPad Prism software, the horizontal coordinate is the antibody concentration and the vertical coordinate is the inhibition rate. The results are shown in FIG. 6A and Table 3. EGFR-HPA8 showed better growth inhibition of MDA-MB-468 cells than SCT200 and Cetuximab control antibodies under ligand-free conditions. EGFR-HPA8 had a similar maximum inhibition rate as SCT200 but had a lower growth inhibition EC₅₀. The results of Example 2.2.1 show that the ability of EGFR-HPA8 to block the binding of EGF-Fc protein to the EGFR on MDA-MB-468 cells was stronger than SCT200 and Erbitux. In this example different ligands of EGFR were added in the MDA-MB-468 cell growth inhibition assay to further validate the ability of EGFR-HPA8 to block EGFR ligand-receptor binding at the cellular functional level. The results are shown in FIG. 6B-6D and Table 3. EGFR-HPA8 inhibited the proliferation of MDA-MB-468 cells better than SCT200 and Cetuximab under different ligand conditions.

TABLE 3 EC50 and maximum neutralization rate of EGFR-HPA8 antibody to inhibit the proliferation of MDA-MB-468 cells Maximum inhibition Ligand Antibody EC₅₀ (nM) rate (%) None EGFR-HPA8 0.90 50.1 SCT200 1.97 52.3 Cetuximab 1.59 44.4 8 ng/mL HB-EGF EGFR-HPA8 2.64 47.8 SCT200 7.52 53.4 Cetuximab 19.9  42.7 200 ng/mL BTC EGFR-HPA8 / 87.5 SCT200 / 40.9 Cetuximab / 20.2 1 μg/mL EREG EGFR-HPA8 0.81 65.2 SCT200 1.44 66.7 Cetuximab 1.47 59.9

2.2.2.2 Ability of Humanized Antibodies to Inhibit Fadu Cell Proliferation

Fadu is a human pharyngeal squamous carcinoma cell strain which highly expresses EGFR and also autocrines various ligand factors of EGFR. Referring to Example 2.2.2.1, inhibition of proliferation of Fadu cells (source: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) by EGFR-HPA8 antibody under different ligand conditions was measured by WST-8 assay. The results are shown in FIG. 7 and Table 4. The growth inhibitory ability of EGFR-HPA8 on Fadu cells was similar to that of SCT200 and better than that of Cetuximab under the ligand-free condition, while the growth inhibitory ability of EGFR-HPA8 on Fadu cells was significantly better than that of SCT200 and Cetuximab under different ligand conditions. The results showed that the EGFR-HPA8 antibody had a better ability to inhibit EGFR ligand-receptor binding compared with SCT200 and Cetuximab.

TABLE 4 EC50 and maximum neutralization rate EGFR-HPA8 antibody to inhibit the proliferation of Fadu cells Maximum inhibition Ligand Antibody EC₅₀ (nM) rate (%) None EGFR-HPA8 0.126 47.6 SCT200 0.129 49.7 Cetuximab 0.251 46.0 4 ng/mL EGF EGFR-HPA8 / 74.4 SCT200 / 68.5 Cetuximab / / 8 ng/mL HB-EGF EGFR-HPA8 / 55.8 SCT200 / 65.0 Cetuximab / 13.6 200 ng/mL BTC EGFR-HPA8 / 49.5 SCT200 / / Cetuximab / / 1 μg /mL EREG EGFR-HPA8 10.9 48.4 SCT200 14.5 48.3 Cetuximab 17.7 44.8

2.2.3 Humanized Antibody ADCC Effects

This example uses a recombinant CD16a reporter gene system to detect the humanized antibody EGFR-HPA8-mediated ADCC effects. The recombinant CD16a reporter gene system includes effector cells Jurkat-NFAT-Luc2p-CD16A and target cells expressing EGFR. When the two cells are co-cultured and the EGFR antibody is added simultaneously, the Fab fragment of the EGFR antibody binds to EGFR expressed on the surface of the target cells, and its Fc fragment can bind to the effector cells overexpressing Fcγ receptor CD16a thereby activating effector cells Jurkat-NFAT-Luc2p-CD16A and promoting NFAT-RE-mediated bioluminescence.

The target cells A431 cells (source: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were uniformly inoculated in 96-well plates at 1×10⁴/well. After overnight incubation, different concentrations of antibodies (2.67 nM, 0.53 nM, 0.11 nM, 0.021 nM, 0.0043 nM, 0.00085 nM, 0.00017 nM and 0.000034 nM) were added at 40 L/well, followed by 1×10⁵ effector cells Jurkat-NFAT-Luc2p-CD16A (Source: Sinocelltech Limited, same as below), 40 μL/well, 3 replicate wells were used for each assay. Target cells, effector cells, and negative antibody control wells were also used. Cells were incubated for 4 h in a CO₂ incubator at 37° C. and 5% CO₂, and Passive Lysis 5× Buffer, 20 μL/well, was added. The cells were freeze-thawed once, and 20 μL of supernatant per well was transferred to a 96-well white-bottom plate after shaking the plate and mixing, and the luminescence detection was performed by LB960-Microplate Luminol Detector. The quantitative efficacy curves were analyzed and plotted using GraphPad Prism software, with the horizontal coordinate being the concentration of the sample and the vertical coordinate being the RLU value. The bioluminescence intensity induction multiplier=sample RLU/negative control RLU. The results are shown in FIG. 8 , EGFR-HPA8, positive controls Erbitux and SCT200 can mediate effective ADCC effects on EGFR-expressing A431 tumor cells. Among them, EGFR-HPA8 had an advantage in the half effective concentration of the effect and the induction fold, with an EC₅₀ of 0.008 nM and R²=0.999 for the induction of ADCC.

2.3 Efficacy of EGFR-HPA-8 on Human Gastric Cancer Cell Line SNU-5 and Human Non-Small Cell Lung Cancer Line NCI-H1975 Subcutaneously Xenografted Tumors in Mice

Logarithmic growth stage SNU-5 cells (ATCC cell bank) were washed with PBS and digested with 0.25% trypsin, and the digest was collected followed by centrifugation at 800 r/min for 5 min. The cells were resuspended in PBS and the cell concentration was adjusted to 5.0×10⁷ cells/mL (with 50% matrix gel). Balb/c-nude mice (Beijing Viton Lever Laboratory Animal Technology Co.) were subcutaneously inoculated with 5.0×10⁶ SNU-5 cell suspensions on the right dorsum, 100 μL/per mouse. After the tumor volume reached about 170 mm³, the mice were randomly divided into seven groups according to the tumor volume, with five mice in each group. The medicament was administered by intraperitoneal injection (I.P.) on the day of grouping and was given twice a week for 7 consecutive doses. The specific dosing regimen is shown in Table 5 below.

TABLE 5 Trail grouping and dosing Number of Subject Administration Group animals medicament Dose(mg/kg) Administration route frequency 1 4 Solvent control / Intraperitoneal injection BIW 2 5 SCT200 5 Intraperitoneal injection BIW 3 5 SCT200 20 Intraperitoneal injection BIW 4 5 EGFR-HPA8 5 Intraperitoneal injection BIW 5 5 EGFR-HPA8 20 Intraperitoneal injection BIW Note: The administration volume was calculated based on the body weight of mice at 10 mL/kg.

Tumor Growth Inhibition Value (TGI) was calculated as follows: T/C (%)=T_(RTV)/C_(RTV)×100% (T_(RTV) is RTV of treatment group; C_(RTV) is RTV of the negative control group), relative tumor volume RTV=V_(T)/V₀, V₀ is the tumor volume measured by D₀, the grouping and starting dosing day, V_(T) is the tumor volume measured each time. TGI (%)=1−T/C (%).

All experimental animals were in good condition and showed some increase in body weight throughout the course of administration. There was no significant difference in the body weight of mice in each dosing group compared to the solvent control (P>0.05). The changes in body weights of all animals are shown in FIG. 9 and Table 6.

TABLE 6 Effect of EGFR-HPA8 on body weight in SNU-5 human gastric cancer xenografted tumor model mice Weight change Body weight (g) thoroughout after the course of Subject Does Number of Before 31 days of P administration Group medicament (mg/kg) animals administration ^(a) administration value^(b) (g) 1 Solvent / 4 19.3 ± 0.5 20.1 ± 0.7 / 0.78 control 2 SCT200 5 5 18.2 ± 0.7 19.6 ± 1.1 0.6 1.42 3 SCT200 20 5 18.4 ± 0.9 19.5 ± 0.9 0.4 1.16 4 EGFR- 5 5 18.7 ± 1.2 20.2 ± 1.0 0.9 1.44 HPA8 5 EGFR- 20 5 18.8 ± 1.2 19.6 ± 0.6 0.4 0.78 HPA8 ^(a) Mean ± standard deviation ^(b)Statistical comparison of body weight in the treatment group with that in the solvent control group after 31 days of administration, t-test.

The tumor volume and TGI results for each group in the trial are shown in Table 7 and FIG. 10 . After 31 days of group dosing, the mean tumor volume in the Vehicle group was 559.9±144.9 mm³ and in the positive control SCT200 low-dose 5 mpk group was 317.1±197.6 mm³ with a TGI of 44.3%, which was not significantly different from the Vehicle group (P=11.28%). The EGFR-HPA8 low dose 5 mpk group showed better efficacy with a tumor volume of 113.8±74.0 mm³ and TGI of 80.0%, a significant difference compared to the Vehicle group (P<0.05), indicating that EGFR-HPA8 showed slightly better tumor-inhibiting properties than the positive control SCT200 at this dose (P=0.09). The tumor volume in the positive control SCT200 20 mpk group was 191.8±189.1 mm³ with a TGI of 66.5%, which was significantly different from the Vehicle group (P<0.05). The tumor volume in the EGFR-HPA8 high dose 20 mpk group was 175.0±175.0 mm³ with a TGI of 68.9%, which was significantly different from the Vehicle group (P<0.05). EGFR-HPA8 and positive control SCT200 20 mpk group had a significant tumor-inhibiting effect, and there was no significant difference in tumor volume between the two administration groups (P=0.9). In conclusion, EGFR-HPA8 molecules showed significant antitumor efficacy in SNU-5 human gastric cancer xenografted tumor model at both 5 mpk and 20 mpk dose levels, and the tumor-inhibiting effect was slightly better than that of SCT200 at low doses.

TABLE 7 Effect of EGFR-HPA8 on tumor volume in SNU-5 human gastric cancer xenografted tumor model mice Tumor volume of SCT200 5 mpk SCT200 20 mpk EGFR-HPA8 5 mpk EGFR-HPA8 20 mpk solvent Tumor TGI Tumor TGI Tumor TGI Tumor TGI Day control volume ^(a) (%) volume (%) volume (%) volume (%) 1 172.8 ± 41.8 170.7 ± 38.3 / 170.3 ± 39.6 / 171.0 ± 35.8 / 168.5 ± 29.6 / 4 168.7 ± 24.7 159.5 ± 52.0 4.3 166.1 ± 33.8 0.1 150.3 ± 30.5 9.9 179.5 ± 38.6 −9.1 8 249.9 ± 34.0 185.1 ± 66.4 25 186.7 ± 45.0 24.2 139.7 ± 33.6 43.5 136.3 ± 47.2 44 11 263.8 ± 30.8 191.0 ± 61.8 26.7 160.5 ± 53.4 38.3 140.8 ± 37.5 46.1 122.5 ± 69.6 52.4 15 307.3 ± 49.2 185.4 ± 44.2 38.9 192.9 ± 68.9 36.3 140.5 ± 40.3 53.8 146.9 ± 84.5 51 18 330.8 ± 45.5 214.4 ± 57.0 34.4  171.4 ± 106.5 47.5 140.6 ± 36.3 57 156.2 ± 88.8 51.5 22 365.2 ± 78.5 211.3 ± 96.3 41.3  145.5 ± 101.6 59.5  90.8 ± 80.0 74.8 105.4 ± 91.4 70.4 31  559.9 ± 144.9  317.1 ± 197.6 44.3  191.8 ± 189.1 66.5 113.8 ± 74.0 80  175.0 ± 175.0 68.9 ^(a) Mean ± standard deviation

EGFR-HPA8 also showed significant tumor-inhibiting effects on the tumor of human non-small cell lung cancer cell line NCI-H1975 subcutaneous xenografted mouse model. Balb/c-nude mice were subcutaneously inoculated with NCI-H1975 cells on the right dorsum (Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences), and the Grouping and Dosing are shown in Table 8.

TABLE 8 Grouping and Dosing Number of Subject the course of Group animals medicament Dose(mg/kg) Administration route administration 1 4 Solvent control / Intraperitoneal injection BIW × 3 2 5 SCT200 5 Intraperitoneal injection BIW × 3 3 5 SCT200 20 Intraperitoneal injection BIW × 3 4 5 EGFR-HPA8 5 Intraperitoneal injection BIW × 3 5 5 EGFR-HPA8 20 Intraperitoneal injection BIW × 3 Note: The administration volume was10 mL/kg the body weight of mice.

All experimental animals were in good condition and showed some increase in body weight throughout the course of administration, and there was no significant difference in the body weight of mice in each administration group compared with the solvent control (P>0.05). The changes in body weight of all animals are shown in FIG. 11 and Table 9.

TABLE 9 Effect of EGFR-HPA8 on body weight in NCI-H1975 human lung cancer xenografted tumor model mice Weight change Body weight (g) thoroughout after the course of Subject Does Number of Before 31 days of P administration Group medicament (mg/kg) animals administration ^(a) administration value^(b) (g) 1 Solvent / 6 19.1 ± 1.4 20.3 ± 1.5 / +1.2 control 2 SCT200 5 6 19.2 ± 1.0 20.1 ± 1.2 0.79 +0.9 3 SCT200 20 6 19.9 ± 0.8 21.1 ± 1.0 0.28 +1.2 4 EGFR- 5 6 20.0 ± 1.2 20.7 ± 1.2 0.58 +0.7 HPA8 5 EGFR- 20 6 20.0 ± 1.1 21.8 ± 0.7 0.05 +1.8 HPA8 ^(a) Mean ± standard deviation ^(b)Statistical comparison of body weight in the treatment group with that in the solvent control group after 31 days of administration, t-test.

The tumor volume and TGI results for each group in the trial are shown in Table 10 and FIG. 12 . After 18 days of grouping and dosing, the positive control SCT200 and EGFR-HPA8 high and low dose groups showed significant tumor-inhibiting effects and statistically significant differences in tumor volume between each administration group and the Vehicle group. The mean tumor volume in the Vehicle group on Day 18 was 1564.3±529.0 mm³. The tumor volumes after SCT200 5 mpk and EGFR-HPA8 5 mpk treatment were 151.9±99.1 mm³ and 86.5±107.5 mm³, with TGI of 90.4% and 94.5%, respectively. The tumor volumes after SCT200 20 mpk and EGFR-HPA8 20 mpk treatment were 289.8±321.4 mm³ and 149.3±94.9 mm³, with TGI of 81.8% and 90.4%, respectively. EGFR-HPA8 at both 5 mpk and 20 mpk doses showed a slightly better tumor-inhibiting effect than positive control SCT200, but there was no statistically significant difference. Taken together, these results suggest that the EGFR-HPA8 molecule has significant antitumor efficacy in the NCI-H1975 human non-small cell lung cancer xenografted tumor model.

TABLE 10 Effect of EGFR-HPA8 on tumor volume and TGI in NCI-H1975 human lung cancer xenografted tumor model Tumor volume SCT200 5 mpk SCT200 20 mpk EGFR-HPA8 5 mpk EGFR-HPA8 20 mpk of solvent Tumor TGI Tumor TGI Tumor TGI Tumor TGI Day control volume (%) volume (%) volume (%) volume (%) 1 178.5 ± 79.6 181.0 ± 90.1 / 181.7 ± 86.0  / 179.8 ± 76.1  / 178.0 ± 70.3 / 5  384.6 ± 156.5 173.4 ± 82.2 55.5 242.0 ± 201.2 38.2 119.3 ± 75.1  69.2 150.5 ± 54.1 60.8 8  617.6 ± 219.5 130.5 ± 65.1 79.2 226.0 ± 220.8 64.1 91.8 ± 72.7 85.3 104.3 ± 46.2 83.1 11  937.7 ± 319.8 133.1 ± 87.1 86 223.7 ± 198.0 76.6 80.9 ± 71.3 91.4  91.5 ± 34.9 90.2 15 1308.8 ± 456.6 121.6 ± 74.8 90.8 234.3 ± 158.3 82.4 109.4 ± 145.3 91.7 113.4 ± 56.0 91.3 18 1564.3 ± 529.0 151.9 ± 99.1 90.4 289.8 ± 321.4 81.8  86.5 ± 107.5 94.5 149.3 ± 94.9 90.4 ^(a)Mean ± standard deviation

Example 3: Design and Test of EGFR Antibody/TGFβR2 Fusion Proteins Containing Different Truncated Forms of TGFβR2

3.1 Design of EGFR Antibody/TGFβR2 Fusion Protein Expression Vectors of Different Truncated Forms of TGFβR2, Protein Expression and Purification

This embodiment uses anti-EGFR antibody as the targeting portion of the fusion protein and TGFβR2 extracellular domain as the immunomodulatory part of the fusion protein. The TGFβR2 extracellular domain is linked to the C-terminal of the heavy chain of the EGFR antibody by homologous recombination, and the EGFR antibody/TGFβR2 extracellular domain fusion protein (EGFR/TGFβR2) is formed by two chains, the light chain, and the heavy chain. The structure of the fusion protein is shown in FIG. 13 . The mass spectrometry results showed that there were multiple susceptibility sites between the N-terminal 7-15 sites of the full-length TGFβR2 extracellular domain. To improve the structural stability of the fusion protein, the N-terminal amino acid sequence of the extracellular domain of TGFβR2 was modified with amino acid deletions in different amino acid residue numbers in this example (SEQ ID NO:47-65). The EGFR antibody/TGFβR2 fusion protein links the amino acid of the heavy chain C-terminal of the EGFR antibody to the extracellular domain of TGFβR2 with different amino acid deletion forms via the (G₄S)₄ Linker (SEQ ID NO:66). In addition, the heavy chain C-terminal lysine of the EGFR antibody was removed to reduce the risk of proteolysis. The specific protocol of the EGFR antibody/TGFβR2 fusion protein design are shown in Table 11.

TABLE 11 EGFR antibody/TGFβR2 fusion protein protocol TGFβR2 Sequence description Number of N- extracellular of the TGFβR2 terminal amino domain Heavy chain Samples extracellular domain acid deletions SEQ ID SEQ ID Fusion protein1 Full length / SEQ ID NO: 47 SEQ ID NO: 88 Fusion protein2 ECD delete (6-16) 11 SEQ ID NO: 48 SEQ ID NO: 89 Fusion protein3 ECD delete (6-19) 14 SEQ ID NO: 49 SEQ ID NO: 90 Fusion protein4 ECD delete (6-21) 16 SEQ ID NO: 50 SEQ ID NO: 91 Fusion protein5 ECD delete (6-25) 20 SEQ ID NO: 51 SEQ ID NO: 92 Fusion protein6 ECD delete (6-26) 21 SEQ ID NO: 52 SEQ ID NO: 93 Fusion protein7 ECD delete (6-27) 22 SEQ ID NO: 53 SEQ ID NO: 94 Fusion protein8 ECD delete (7-26) 20 SEQ ID NO: 54 SEQ ID NO: 95 Fusion protein9 ECD delete (5-26) 22 SEQ ID NO: 55 SEQ ID NO: 96 Fusion protein10 ECD delete (4-26) 23 SEQ ID NO: 56 SEQ ID NO: 97 Fusion protein11 ECD delete (3-26) 24 SEQ ID NO: 57 SEQ ID NO: 98 Fusion protein12 ECD delete (2-26) 25 SEQ ID NO: 58 SEQ ID NO: 99 Fusion protein13 ECD delete (1, 6-26) 22 SEQ ID NO: 59 SEQ ID NO: 100 Fusion protein14 ECD delete (1-2, 6-26) 23 SEQ ID NO: 60 SEQ ID NO: 101 Fusion protein15 ECD delete (1-3, 6-26) 24 SEQ ID NO: 61 SEQ ID NO: 102 Fusion protein16 ECD delete (1-4, 6-26) 25 SEQ ID NO: 62 SEQ ID NO: 103 Fusion protein17 ECD delete (1-19) 19 SEQ ID NO: 63 SEQ ID NO: 104 Fusion protein18 ECD delete (1-21) 21 SEQ ID NO: 64 SEQ ID NO: 105 Fusion protein19 ECD delete (1-26) 26 SEQ ID NO: 65 SEQ ID NO: 106

In the above protocol, the target gene was amplified by PCR or Overlap-PCR, ligated to the expression vector by in-fusion, and the plasmids were extracted separately after sequencing verification, and transiently transferred to HEK-293 cells (fut8 knockout) and cultured until day 7. Centrifuge and collect the supernatant. The cell supernatant after centrifugation was purified using Protein A affinity chromatography to obtain ADCC-enhanced EGFR antibody/TGFβR2 fusion protein.

3.2 Degradation of EGFR Antibody/TGFβR2 Fusion Proteins Containing Different Truncated Forms of TGFβR2

The purity as well as the degradation of the expression product was analyzed by reduced SDS-PAGE. The different truncated EGFR antibody/TGFβR2 fusion proteins 1-16 purified in Example 3.1 were subjected to the reduced SDS-PAGE. Specific steps of reduced SDS-PAGE: (1) SDS-PAGE preparation: 3.9% concentrating gel, 13% separating gel; (2) samples were boiled at 100° C. for 2 min, centrifuged, and then sampled with 8 μg; (3) constant current of 40 mA, electrophoresis time of 1 h. The results are shown in FIG. 14 . The molecular weight of the light chain of EGFR antibody/TGFβR2 fusion protein was about 25 KDa, the molecular weight of the heavy chain was about 66 KDa, and the molecular weight of the clipping species was between 45-66 KDa. The results showed that there were obvious clipping species in the extracellular domain of TGFβR2, while the different truncated forms fusion proteins of the TGFβR2 extracellular domain had substantially fewer bands resulted from the degradation than the full-length forms fusion proteins of the TGFβR2 extracellular domain. Thus, the different truncated forms of the TGFβR2 extracellular domain prepared in the present invention significantly enhanced the stability of the TGF-β receptor-containing antibody fusion protein.

3.3 Degradation Tendency of EGFR Antibody/TGFβR2 Fusion Proteins Containing Different Truncated Forms of TGFβR2

The 293E cell supernatant which accelerates fusion proteins processing is used to further evaluate the stability of different truncated forms of EGFR antibody/TGFβR2 fusion proteins. 293E cell expression system, which is often used to express antibodies, expresses a variety of host cell proteins (HCPs) and proteases required for cell growth. Therefore, the stability of antibodies can be assessed by observing the degradation propensity of fusion proteins in the supernatant of 293E cells.

The purified fusion protein was mixed with the supernatant of 293E cells cultured for 10 days at a volume ratio of 1:0.3, and the final concentration of fusion protein was about 1 mg/mL. The mixed samples were shaken and mixed well and incubated at 37° C. for 48 h. A control of incubation without cell supernatant was also used. The purity of the samples and the content of clipping species were detected by reduced SDS-PAGE, and the fusion protein heavy chain purity was calculated by BandScan software.

The results of the assay are shown in FIG. 15 , and the fracture percentage in each sample is shown in Table 12. The results showed that the percentage of sheared bodies of TGFβR2 truncated form fusion protein (fusion protein 2-16) in the control group were all less than 4.0%, which was much lower than that of TGFβR2 full-length form fusion protein (fusion protein 1) at 24.8%. After incubation with the same cell supernatant for 48 h at 37° C., all of the TGFβR2 full-length form fusion proteins (fusion protein 1) were sheared in the experimental group, with a percentage of 100%. The truncated forms of TGFβR2 fusion proteins (fusion proteins 2-16) contained different percentages of clipping species but were significantly better than the full-length control, wherein fusion protein 2, fusion protein 6, fusion protein 9, fusion protein 10, and fusion protein 13 showing the best performance, with the percentage of clipping species content less than 3.0%.

TABLE 12 Degradation ratio of EGFR antibody/TGFβR2 fusion proteins containing different truncated forms of TGFβR2 Experimental group Control group Percentage Percentage Percentage Percentage of heavy of clipping of heavy of clipping Samples chain % species % chain % species % Fusion protein1 0 100 75.2 24.8 Fusion protein2 97.3 2.7 96.8 3.2 Fusion protein3 96.4 3.6 96.1 3.9 Fusion protein4 91.2 8.8 97.6 2.4 Fusion protein5 91.0 9.0 98.1 1.9 Fusion protein6 97.2 2.8 97.2 2.8 Fusion protein7 6.8 93.2 97.2 2.8 Fusion protein8 87.7 12.3 97.5 2.5 Fusion protein9 98.0 2.0 98.6 1.4 Fusion protein10 98.3 1.7 99.0 1.0 Fusion protein11 95.1 4.9 99.2 0.8 Fusion protein12 40.8 59.2 99.7 0.3 Fusion protein13 97.5 2.5 96.8 3.2 Fusion protein14 93.9 6.1 100 0 Fusion protein15 81.6 18.4 99.1 0.9 Fusion protein16 30.0 70.0 99.0 1.0

The above results indicate that the EGFR antibody/TGFβR2 fusion proteins of different truncated forms of TGFβR2 are more resistant to protease degradation than TGFβR2 fusion protein 1 of the full-length form.

3.4 Binding Assay of EGFR Antibody/TGFβR2 Fusion Protein of Different Truncated Forms of TGFβR2 for TGF-β

One hundred ng/mL TGF-β1 and 40 ng/mL EGFR-His proteins were coated on 96-well plates at 100 μL/well, respectively, overnight at 4° C. The plates were washed the next day, blocked at room temperature for 1 h, and 2 μg/mL of EGFR antibody/TGFβR2 fusion protein of different truncated forms of TGFβR2 was added at 100 μL/well. Washed the plate after 1 h incubation to remove unbound antibody, added secondary antibody Goat anti-hIgG Fc/HRP and incubated and repeated the wash. Finally, the substrate chromogenic solution was added for color development, and the OD₄₅₀ was read by the a microplate reader after termination. The results are shown in FIG. 16 . The binding ability of EGFR antibody/TGFβR2 fusion protein with different truncated forms of TGFβR2 to TGF-β1 was different, but the binding ability to EGFR was similar.

3.5 Neutralizing Assay of EGFR Antibody/TGFβR2 Fusion Protein of Different Truncated Forms of TGFβR2 for TGF-β

TGF-β regulates cellular functions by regulating the transcription of multiple target genes.

Plasminogen activator inhibitor 1 (PAI-1) is an important target of the TGF-β1/Smad signaling pathway downstream, and activates Smad3 binds to the cis-acting element of the PAI-1 promoter region to regulate the expression of PAI-1. The element containing the PAI-1 promoter region is inserted in a specific form into a luciferase-containing vector and transferred into HepG2 cells. In this reporter gene system, the addition of exogenous TGF-β protein initiates the expression of the luciferase reporter gene and luminescence in the presence of substrate. When an exogenous TGF-β antibody is added, it neutralizes the TGF-β protein, blocks the binding of TGF-β to TGFβR2, inhibits the downstream signaling pathway, and finally inhibits the expression of the luciferase reporter gene. Therefore, the in vitro efficacy of the TGF-β antibody neutralizing TGF-β can be determined by detecting the intensity of the light signal.

The 96-well plates were uniformly inoculated with HepG2-3TP-Luc2p-puro cells (source: Sinocelltech Limited, same below) at 30,000 cells/well. After overnight monolayer culture, the medium in the 96-well plate was discarded and replaced with a DMEM medium containing 0.5% FBS and incubated for 6 h at 37° C. in a 5% CO₂ incubator. The medium in the 96-well plate was discarded and 4 ng/mL TGF-β1 protein was added, along with EGFR antibody/TGFβR2 fusion protein at a final concentration of 0.02 μg/ml, and incubated for 18 h at 37° C. in a 5% CO₂ incubator. The negative control M group (containing cells and TGF-β1) and the negative control M′ group (containing cells without TGF-β 1) were used at the same time. Finally, 5× Lysis buffer was added and 10 μL of cell samples were taken to detect the bioluminescence intensity value (RLU), and the neutralization rate of the EGFR antibody/TGFβR2 fusion protein was calculated. Neutralization rate (%)=(RLU value of M Group−RLU value of the sample)/(OD value of M Group−OD value of M′ Group)×100%. The concentration of antibodies was used as the horizontal coordinate and the antibody neutralization rate was used as the vertical coordinate, and the quantitative efficacy curves were analyzed and plotted using GraphPad Prism software. As shown in FIG. 17 , EGFR antibody/TGFβR2 fusion protein 2, fusion protein 6, fusion protein 8, fusion protein 13, fusion protein 14, and fusion protein 16 containing the truncated forms of TGFβR2 had a certain ability to neutralize TGF-β 1, among which fusion protein 2 had similar neutralization ability as fusion protein 1 containing the full-length form of TGFβR2, while fusion protein 5, fusion protein 6 and fusion protein 8 showed better neutralization ability than fusion protein 1 at this concentration. Fusion protein 6 exhibited the strongest ability to neutralize TGF-β1. The remaining EGFR antibody/TGFβR2 fusion proteins containing the truncated form of TGFβR2 had essentially no or weak neutralizing ability.

The TGF-β3 protein has a high affinity for TGFβR2 and can activate TGF-β downstream signaling. The ability of the fusion protein to neutralize TGF-β3 (20 ng/mL final concentration) was assayed by applying the reporter gene system in this example. The results are shown in FIG. 18 . Fusion protein 2, fusion protein 4, and fusion protein 13 had a similar ability to neutralize TGF-β3 as fusion protein 1, while fusion protein 3, fusion protein 5, fusion protein 6, and fusion protein 8 had a better ability to neutralize TGF-β3 than fusion protein 1. Fusion protein 6 also showed the strongest ability to neutralize TGF-β3.

Based on the above analysis of the stability and neutralization ability of fusion proteins containing TGFβR2 truncated forms, the present invention prefers TGFβR2 truncated forms of fusion proteins 2˜6, 8, 13, 14, and 16, more preferably TGFβR2 truncated forms of fusion proteins 2, 5, 6 and 8, and most preferably TGFβR2 truncated forms of fusion protein 6.

Example 4: In Vitro Biological Property of Fusion Protein 6, an EGFR Antibody/TGFβR2 Fusion Protein of TGFβR2 Truncated Form

4.1 Assay of Binding Ability of EGFR Antibody/TGFβR2 Fusion Protein

4.1.1 Properties of EGFR Antibody/TGFβR2 Fusion Protein Binding and Competing for TGF-β

TGF-β1 protein and TGF-β3 protein at a final concentration of 2 μg/mL were coated on 96-well plates at 100 μL/well, respectively, and coated overnight at 4° C. The plates were washed the next day, blocked at room temperature for 1 h, and incubated with different concentrations (1.22 pM, 4.88 pM, 19.53 pM, 78.13 pM, 312.5 pM, 1250 pM, 500 pM, 2000 pM) of EGFR antibody/TGFβR2 fusion protein 6 for 1 h. After that, the plates were washed to remove unbound antibodies, and incubated with secondary antibody Goat anti-hIgG₅₀ ² F(ab)2/HRP and were repeatedly washed, the substrate chromogenic solution was added for color development, and the OD₄₅₀ was read by a microplate reader after termination. The results are shown in FIG. 19 . The binding ability of fusion protein 6 for TGF-β1 and TGF-β3 proteins was similar to that of fusion protein 1, with an EC₅₀ of 91 pM and R²=0.998 for binding to TGF-β1 protein, and with an EC₅₀ of 102 pM and R²=0.998 for binding to TGFβ3 protein.

This example further analyzes the ability of fusion protein 6 to compete with TGF-β1 protein or TGF-β3 protein for binding to TGFβR2-Fc protein at the protein level.

TGF-β1 protein at a final concentration of 0.2 μg/mL or TGF-β3 protein at 0.5 μg/mL was coated on a 96-well plate at 100 μL/well and overnight at 4° C. The plates were washed the next day, blocked at room temperature for 1 h, and 100 μL of EGFR antibody/TGFβR2 fusion protein at different concentrations (0.05 nM, 0.14 nM, 0.42 nM, 1.25 nM, 3.75 nM, 11.24 nM, 33.71 nM, 101.12 nM) were added with 100 μL of the final concentration of 0.2 μg/mL (TGF-β1 competition) or 1 μg/mL (TGF-β3 competition) of biotin-labeled TGFβR2-Fc protein (protein source: Sino Biological, Inc. and biotin-labeled by Sinocelltech Limited, same below). Positive control wells with TGFβR2-Fc protein only were also used. The plate was washed after 1 h incubation, and the plate was repeatedly washed after 1 h incubation by adding detection secondary antibody Streptavidin/HRP. Finally, the substrate chromogenic solution was added for color development, and after termination, the a microplate reader read OD₄₅₀. The competitive inhibition rate PI % of the fusion protein was calculated based on the OD₄₅₀ value, and the inhibition rate PI (%)=(OD₄₅₀ value of positive wells−OD₄₅₀ value of sample wells)/OD450 value of positive wells×100%. The results are shown in FIG. 20 , and fusion protein 6 has a similar ability to block the binding of TGF-β1 protein or TGF-β3 protein to TGFβR2-Fc as fusion protein 1.

4.1.2 Binding Properties of EGFR Antibody/TGFβR2 Fusion Protein for EGFR

Referring to Example 1.2.1, the binding ability of the fusion protein to recombinant human EGFR protein was measured by ELISA. As shown in FIG. 21 , the binding ability of fusion protein 6 to EGFR and the ability to compete with EGF to bind to EGFR was similar to that of EGFR-HPA8, with a binding EC₅₀ of 133.1 ng/mL and R²=1.000.

4.2 EGFR Antibody/TGFβR2 Fusion Protein Binding Affinity Assay

In this example, the affinity of EGFR antibody/TGFβR2 fusion protein binding to biotinylated recombinant human EGFR protein and TGF-β1 protein was determined using a biomolecular interaction analysis system (model: OctetRED96e, manufacturer: Fortebio) with EGFR-HPA8 and H7N9-R1, respectively, as negative controls. Affinity parameters were derived by fitting multiple concentration point binding and dissociation curves, and the results are shown in Tables 13 and 14, and the specific kinetic characteristic parameter curves are shown in FIGS. 22 and 23 .

The results showed that fusion protein 6 retained a higher binding affinity to human EGFR protein compared to the monoclonal antibody EGFR-HPA8, with a KD value of 8.77 pM, a binding constant k_(on) value of 1.68E+06 M⁻¹ s⁻¹ and a dissociation constant k_(d) is of 1.47E-05 s⁻¹. Furthermore, the truncated form of TGFβR2 fusion protein 6 has a similar affinity to human EGFR protein to the full-length form of TGFβR2 fusion protein 1 with a k_(D) value of 96.1 pM, a binding constant k_(on) value of 1.53E+06 M s⁻¹⁻¹, and a dissociation constant k_(d) is of 1.47E-04 s.⁻¹

TABLE 13 Affinity of truncated EGFR antibody/TGFβR2 fusion protein to recombinant human EGFR protein Sample K_(D) (M) kon (M⁻¹s⁻¹) kdis (s⁻¹) EGFR-HPA8 6.90E−12 1.59E+06 1.10E−05 Fusion protein 6 8.77E−12 1.68E+06 1.47E−05

TABLE 14 Affinity of truncated EGFR antibody/TGFβR2 fusion protein to recombinant human TGF-β1 protein Sample K_(D) (M) kon (M⁻¹s⁻¹) kdis (s⁻¹) Fusion protein 1 1.73E−10 1.45E+06 2.51E−04 Fusion protein 6 9.61E−11 1.53E+06 1.47E−04

The above results indicate that fusion protein 6 has a good affinity with both human EGFR and TGF-β 1.

4.3 Assay of EGFR Antibody/TGFβR2 Fusion Protein Neutralizing TGF-β

TGF-β1 can inhibit the proliferation of Mv-1-lu cells, so the ability of EGFR antibody/TGFβR2 fusion protein to neutralize TGF-β1 can be detected using the WST-8 assay.

Mv-1-lu cells (source: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were inoculated uniformly with 50 μL/well in a 96-well plate at a cell inoculation density of 1×10³/well. Cells were incubated in a CO₂ incubator for about 3 h to adhere to the well wall, and then EGFR antibody/TGFβR2 fusion protein samples at different concentrations (0.0078 nM, 0.0156 nM, 0.0313 nM, 0.0625 nM, 0.125 nM, 0.25 nM, 0.5 nM, 1 nM, 2 nM) diluted in 1640 medium containing 10% FBS was added with 50 μL/well. Finally, the TGF-β1 factor at a final concentration of 1 ng/mL was added with 10 μL/well. Positive control M group (containing cells and TGF-β1), negative control M′ group (containing cells without TGF-β1), and blank control B group (adding only culture medium without cells) were also used. Cells were incubated in CO₂ incubator at 37° C. and 5% CO₂ for 5 days and then WST-8 was added at 10 μL/well. The samples were left for 180 min and the reading of OD₄₅₀-OD₆₃₀ was measured by a microplate reader, and the neutralization rate of EGFR antibody/TGFβR2 fusion protein was calculated by subtracting the reading of blank control B. Neutralization rate (%)=(OD of M′ group−OD of the sample)/(OD of M′ group−OD of M group)×100%, and the quantitative efficacy curve was analyzed and plotted using GraphPad Prism software, the horizontal coordinate is the antibody concentration and the vertical coordinate is the inhibition rate. As shown in FIG. 24 , both fusion protein 6 and the control product H7N9-R1-43-IgG1 (L9) of TGFβR2 (source: Sinocelltech Limited for biotin labeling, same below) could effectively neutralize the inhibition of Mv-1-lu proliferation by TGF-β1 in a dose-dependent relationship, and the half effective concentration of fusion protein 6 was smaller than that of the control product H7N9-R1-43-IgG1 (L9), indicating a better neutralizing activity of the molecule. EGFR-HPA8 did not neutralize TGF-β1, suggesting that it is the TGFβR2 portion of fusion protein 6 that shows a neutralizing effect on TGF-β1.

The ability of fusion protein 6 to neutralize TGF-β was further assayed by using the reporter gene system in this example, referring to Example 3.5. The results are shown in FIG. 25 , and both fusion protein 6 and the control product H7N9-R1-43-IgG1 (L9) of TGFβR2 can effectively neutralize TGF-β1 in a dose-dependent relationship, and the maximum neutralization rate of fusion protein 6 (74.8%) was much higher than that of the control product H7N9-R1-43-IgG1 (L9) (55.3%), further indicating the excellent neutralizing activity of fusion protein 6.

4.4 Assay of Cell Proliferation Inhibitory Activity of EGFR Antibody/TGFβR2 Fusion Protein

The EGFR antibody/TGFβR2 fusion protein inhibition on the growth of MDA-MB-468 cells was assayed by the WST-8 method according to Example 1.2.2 to determine its EGFR portion property. As shown in FIG. 26 , the ability of fusion protein 6 to inhibit the proliferation of MDA-MB-468 cells was similar to that of EGFR-HPA8, and the inhibition rate increased with the increase of medicament concentration in an “S” curve. The control product H7N9-R1-43-IgG1 (L9) with TGFβR2 function did not inhibit the proliferation of MDA-MB-468 cells, indicating that fusion protein 6 inhibited the proliferation of tumor cells by its EGFR portion.

4.5 ADCC Effect of EGFR Antibody/TGFβR2 Fusion Protein

The ADCC effect mediated by EGFR antibody/TGFβR2 fusion protein on EGFR-expressing cells was assayed with reference to Example 2.2.3. The results are shown in FIG. 27 . In the concentration range of 0.00004-3 nM, fusion protein 6 and anti-EGFR antibody EGFR-HPA8 can mediate similar ADCC effects on EGFR-expressing tumor cells A431. The control product H7N9-R1-43-IgG1 (L9) with TGFβR2 function had no ADCC effect on A431 cells, indicating that it is the EGFR portion of fusion protein 6 that mediates the ADCC function in this experimental system.

Example 5: Pharmacodynamic Study on TGFβR2 Truncated Form (Fusion Protein 6) EGFR Antibody/TGFβR2 Fusion Protein in NCI-H1975 Subcutaneous Graft Tumor Model

Balb/c-nu mice were subcutaneously inoculated with 1×10⁶ NCI-H1975 cells on the right side of the rib cage. When the tumor volume reached about 300 mm³, the animals were randomly grouped by tumor volume to 6 animals in each group and administered. Dosing was started on the day of the grouping, the medicament was administered by intraperitoneal injection (I.P.) twice a week for 10 consecutive doses, and was discontinued after the last dose to observe tumor recurrence. The specific dosing regimen is shown in Table 15 below.

TABLE 15 Grouping and Dosing Number of Subject the course of Group animals medicament Dose(mg/kg) Administration route administration 1 6 Vehicle / Intraperitoneal injection BIW × 5 w 2 6 EGFR-HPA8-Ae0 20  Intraperitoneal injection BIW × 5 w 3 6 Fusion protein 6 24^(b) Intraperitoneal injection BIW × 5 w Note: The administration volume was10 mL/kg the body weight of mice.

The animals in each group were in a good general condition such as activity and feeding during the course of administration, and their body weight increased to some extent. There was no significant difference in body weight between the dosing group and the solvent control group after dosing (P>0.05). The changes in body weights of all animals are shown in Table 16 and FIG. 28 .

TABLE 16 Effect of fusion protein 6 on the body weight of H1975 non- small cell lung cancer subcutaneous xenografted tumor mice Weight change Body weight (g) thoroughout after the course of Subject Number of Before 35 days of P administration Group medicament animals administration ^(a) administration value^(b) (g) 1 Solvent control 6 18.8 ± 0.2 23.7 ± 0.6 / +4.3 2 EGFR-HPA8 6 18.5 ± 0.5 22.0 ± 0.9 0.164 +2.9 3 Fusion protein 6 6 19.2 ± 0.4 21.9 ± 0.4 0.030 +2.4 ^(a) Means ± standard deviations. ^(b)Statistical comparison of body weight in the treatment group with that in the solvent control group after 35 days of administration, t-test.

The tumor volume results for each group in the trial are shown in Table 17 and FIG. 29 . After 35 days of treatment in groups, the mean tumor volume in the solvent control group was 7150.78±780.4 mm³. Five of the six mice in the fusion protein 6 administration group had complete tumor disappearance (CR) with a mean tumor volume of 4.55±4.55 mm³ and a TGI of 99.9%, which was significantly different from that of the solvent control group (P<0.001). In contrast, only one mouse in the EGFR-HPA8 administration group showed complete tumor disappearance, with a mean tumor volume of 79.44±28.65 mm³ and a TGI of 98.9%. The results indicated that fusion protein 6 had a significant inhibitory effect on NCI-H1975 non-small cell lung cancer subcutaneous transplanted tumors and the tumor-inhibiting effect was superior to that of EGFR-HPA8 at the same molar dose (P=0.237).

TABLE 17 Effect of fusion protein 6 on tumor volume in mice in H1975 non-small cell lung cancer xenografted tumor model Tumor Tumor volume of Tumor volume of solvent volume of fusion control EGFR-HPA8 TGI protein TGI Day (mm³) (mm³) (%) P^(a) 6 (mm³) (%) P^(b) 1 324.77 ± 37.33 323.06 ± 34.66 — 0.9739 323.20 ± 30.48  — 0.9747 4 1004.24 ± 100.19 479.20 ± 57.44 52.00% 0.0011 490.36 ± 38.50  50.90% 0.0007 7 1506.18 ± 138.21 416.80 ± 57.56 72.20% 0 528.37 ± 88.27  64.70% 0.0001 11 2167.62 ± 204.64 425.54 ± 82.69 80.30% 0 288.30 ± 78.16  86.60% 0 14 2825.99 ± 217.47 299.87 ± 60.52 89.30% 0 143.14 ± 47.80  94.90% 0 18 3685.00 ± 626.56 152.98 ± 48.45 95.80% 0 60.65 ± 24.72 98.30% 0 21 4315.02 ± 336.11 118.78 ± 33.71 97.20% 0 37.65 ± 11.93 99.10% 0 25 5031.82 ± 414.57  64.63 ± 16.11 98.70% 0 10.36 ± 4.92  99.80% 0 28 5895.36 ± 504.19  77.86 ± 18.00 98.70% 0 3.86 ± 3.86 99.90% 0 32 6722.83 ± 477.48  85.01 ± 27.00 98.70% 0 2.33 ± 2.33 100.00% 0 35 7150.78 ± 780.4   79.44 ± 28.65 98.90% 0 4.55 ± 4.55 99.90% 0 39  7959.47 ± 1094.45 112.85 ± 39.69 98.60% 0 6.53 ± 6.53 99.90% 0 42  7862.70 ± 1213.66 161.12 ± 59.62 97.90% 0.0001 5.39 ± 5.39 99.90% 0.0001 47  8411.14 ± 1224.57 231.39 ± 90.64 97.20% 0.0001 5.26 ± 5.26 99.90% 0 ^(a)Statistical comparison of tumor volumes in the EGFR-HPA8-Ae0 treatment group with those in the solvent control group, t-test. ^(b)Statistical comparison of tumor volumes in the fusion protein 6 treatment group with those in the solvent control group, t-test.

Example 6: Stability Analysis of TGFβR2 Truncated Form (Fusion Protein 6) EGFR Antibody/TGFβR2 Fusion Protein

6.1 Ultrafiltration Stability Analysis of EGFR Antibody/TGFβR2 Fusion Protein

EGFR antibody/TGFβR2 fusion protein samples were concentrated to a concentration of about 10 mg/mL using ultrafiltration in 100 mM Glycine, 10 mM NaCl, 50 mM Tris, pH 7.5 buffer. The concentrated samples were tested for purity and stability of the ultrafiltered samples by reduced SDS-PAGE and molecular sieve chromatography (SEC-HPLC, Agilent 1260 liquid chromatography system, TSK-G3000SWXL column). EC-HPLC operation steps: (1) mobile phase: 200 mM NaH₂PO₄, 100 mM Arginine, pH 6.5; (2) the loading volume was 80 μg; (3) the analysis time was 30 min, the flow rate was 0.5 m/min, and the column temperature was 25° C.; (4) the purity was calculated according to the normalization method of peak area.

The results of the purity test after sample concentration are shown in FIG. 30 , and the sample purity and fragment ratios are shown in Table 18. The results showed that the TGFβR2 truncated fusion protein 6 was less likely to break after concentration and had higher ultrafiltration stability compared with the full-length TGFβR2 fusion protein.

TABLE 18 SEC results of ultrafiltration stability of EGFR antibody/TGFβR2 fusion protein Reduced SEC (%) SDS-PAGE Fragment Heavy chain Sample Monomers Fragments increase % (%) Fusion protein 1 68.7 24.8 24.8 92.3 Fusion protein 6 99.8 0.1 0.1 95.6

6.2 Thermal Stability Analysis of EGFR Antibody/TGFβR2 Fusion Protein

The thermal stability of the samples was measured by differential scanning fluorimetry (DSF) using an UNcle system (Unchained Labs, model: UNCLE-0330). Operation steps: (1) The sample volume was 9 μL; (2) The experimental parameters were set: the temperature range was 25° C. to 95° C., and the heating rate was 0.3° C./min; (3) the UNcle Analysis software was applied to analyze the data, the midpoint value of the internal fluorescence change curve under UV266 was taken as Tm, and the aggregation onset temperature of the aggregation change curve formed by the static light scattering signal under UV266/Blue473 was taken as Tagg266 and Tagg473.

The results of the thermal stability assay of fusion protein 6 were shown in Table 19, which exhibited good thermal stability.

TABLE 19 Tm assay results of fusion protein 6 Sample Tm (CH2) Tm (Fab) Tagg266 Tagg473 Fusion protein 6 69.7 80.8 73 74.7

6.3 Thermal Accelerated Stability Analysis of EGFR Antibody/TGFβR2 Fusion Protein

After the samples were stored at 45° C. for 1 week, the accelerated stability of the samples was analyzed by SEC-HPLC and SDS-PAGE, and the procedure was the same as 6.1.

The results of thermal accelerated stability of fusion protein 6 are shown in Table 20. After 1 week of storage at 45° C., the SEC purity of fusion protein 6 decreased by 0.7%, but the purity was still high, the level of aggregates increased less and the level of fragments did not change, which showed good thermal accelerated stability.

TABLE 20 Thermal acceleration stability assay results of fusion protein 6 SEC (%) Sample Treatment condition Aggregates Monomers Fragments Fusion No treatment 0 100 0 protein 6 1-week storage at 0.6 99.3 0 45° C. Range of variation 0.6 0.7 0

6.4 Freeze-Thaw Stability Analysis of EGFR Antibody/TGFβR2 Fusion Protein

The samples were stored at −80° C. for 3 h and then transferred to 45° C. for 1 h to thaw, and so on for five repeated freeze-thaws. The freeze-thaw stability of the samples was analyzed by SEC-HPLC, and the procedure was the same as in 6.1.

The results of freeze-thaw stability of fusion protein 6 are shown in Table 21. After five repeated freeze-thaws, the SEC purity of fusion protein 6 did not change significantly, and the levels of aggregates and fragments did not increase significantly, which showed good freeze-thaw stability.

TABLE 21 Freeze-thaw stability assay results of fusion protein 6 SEC (%) Sample Treatment condition Aggregates Monomers Fragments Fusion No treatment 0.1 99.8 0.1 protein 6 repeated freeze-thaws 0.2 99.6 0.2 Range of variation 0.1 0.2 0.1

6.5 Shaking Stability of EGFR Antibody/TGFβR2 Fusion Protein

The samples were placed in deep-well plates and shaken on a vortex shaker at 800 rpm for 24 h. The samples were analyzed by SEC-HPLC with the same procedure as in 6.1. The results are shown in Table 22, which indicates that the SEC monomer purity of the samples did not change significantly after 24 h of shaking, and the levels of aggregates and fragments did not increase significantly, indicating that fusion protein 6 has good shaking stability.

TABLE 22 Shaking stability assay results of fusion protein 6 SEC (%) Sample Treatment condition Aggregates Monomers Fragments Fusion T0 0.3 99.7 0 protein 6 Shaking 0.3 99.7 0

Example 7: Design and Property Assay of Solid Tumor Antigen-Targeting Antibody X/TGFβR2 Fusion Protein, Wherein the TGFβR2 Portion is Truncated Forms of TGFβR2 (Fusion Protein 6)

7.1 Design of TGFβR2 X Antibody/Truncated TGFβR2 Fusion Protein Expression Vector, Expression, and Purification

To further validate the structural stability and the ability to neutralize TGF-β1 of the elected TGFβR2 truncated form in Example 3.2, this Example uses a variety of solid tumor antigens as the targeting portion of the fusion protein and the TGFβR2 extracellular domain (full-length and ECD delete (6-26) fusion protein 6) as the immunomodulatory portion of the fusion protein to form the X antibody/TGFβR2 extracellular domain fusion protein (X/TGFβR2 fusion protein). Similarly, in the X/TGFβR2 fusion protein, the C-terminal amino acid of the X antibody heavy chain links to the TGFβR2 extracellular domain via the (G₄S)₄ Linker. In addition, the C-terminal lysine of the X antibody heavy chain was removed to reduce the risk of proteolytic cleavage. The construction protocol of the X/TGFβR2 fusion protein is shown in Table 23.

TABLE 23 X/TGFβR2 fusion protein design protocol Sample Targeting antibody TGFβR2 ECD Heavy chain SEQ ID Fusion protein 20 Trastuzumab Full length SEQ ID NO: 127 Fusion protein 21 Trastuzumab ECD delete (6-26) SEQ ID NO: 128 Fusion protein 22 Bevacizumab Full length SEQ ID NO: 130 Fusion protein 23 Bevacizumab ECD delete (6-26) SEQ ID NO: 131 Fusion protein 24 Ramucirumab Full length SEQ ID NO: 133 Fusion protein 25 Ramucirumab ECD delete (6-26) SEQ ID NO: 134 Fusion protein 26 Ipilimumab Full length SEQ ID NO: 136 Fusion protein 27 Ipilimumab ECD delete (6-26) SEQ ID NO: 137 Fusion protein 28 Panitumumab Full length SEQ ID NO: 139 Fusion protein 29 Panitumumab ECD delete (6-26) SEQ ID NO: 140

PCR or Overlap-PCR amplifies the target gene and ligates it to the expression vector by in-fusion. The recombinant expression vector was validated by sequencing and the plasmids were extracted, transiently transferred to HEK-293 cells (fut8 knockout), cultured for 7 days and the supernatant was collected by centrifugation. The obtained cell supernatant was purified using Protein A affinity chromatography to purify the fusion protein.

7.2 Degradation of X Antibody/Truncated TGFβR2 Fusion Proteins

The purity, as well as the degradation of the X/TGFβR2 fusion protein, was assayed by reduced SDS-PAGE. The results are shown in FIG. 31 . For the different X/TGFβR2 fusion proteins, the expressed samples of the elected truncated form of TGFβR2 extracellular domain were significantly more stable and had fewer degradation bands than the full-length TGFβR2 extracellular domain control samples. The stability of the samples should be attributed to the present truncated forms of TGFβR2 extracellular domain, no matter what kinds of the targeting portion, i.e. the antibody is used.

7.3 Degradation Tendency of X Antibody/Truncated TGFβR2 Fusion Protein

In this example, 293E cell supernatant was used for the accelerated treatment of the fusion protein, and the degradation stability of X/TGFβR2 fusion protein with the elected truncated TGFβR2 extracellular domain was further determined as in Example 3.3. The results of the sample purity assay are shown in FIG. 32 , and the sample purity and the percentage of shearers are shown in Table 24. The results indicate that X/TGFβR2 fusion proteins with preferred TGFβR2 extracellular domain truncated form is more resistant to protease degradation than the full-length form of TGFβR2 fusion protein.

TABLE 24 Degradation percentage of X/TGFβR2 fusion proteins with truncated TGFβR2 Experimental group Control group Percentage Percentage Percentage Percentage of heavy of clipping of heavy of clipping Samples chain % species % chain % species % Fusion protein 20 15.2 84.8 94.4 5.6 Fusion protein 21 98.1 1.9 96.7 3.3 Fusion protein 22 4.9 94.9 80 20 Fusion protein 23 99.8 0.2 96.2 3.8 Fusion protein 24 24.7 74.8 70.7 29.3 Fusion protein 25 96.8 3.2 96.9 3.1 Fusion protein 26 85.4 14.6 84.4 15.6 Fusion protein 27 97.2 2.8 97.1 2.9 Fusion protein 28 0 100 38.8 61.2 Fusion protein 29 97.1 2.9 98.5 98.5

To assay the stability of the X/TGFβR2 fusion protein with the elected truncated TGFβR2 extracellular domain under certain concentration conditions, its ultrafiltration stability was analyzed using the method of Example 6.1. The results are shown in Table 25. The X/TGFβR2 fusion protein with the preferred truncated extracellular domain of TGFβR2 is less prone to degradation after concentration, and the percentage of clipping species content is less than 4.0% (SDS-PAGE purity), which has stronger ultrafiltration stability than the X/TGFβR2 fusion protein with the full-length extracellular domain of TGFβR2.

TABLE 25 SEC and SDS degradation assay of concentrated X/TGFβR2 fusion protein Reduced SEC-HPLC SDS-PAGE Intact Fragments Fragments clipping Fusion protein No. protein % % increase % species (%) Fusion protein 20 93.1 1.5 −1.1 19.8 Fusion protein 21 94.3 0.5 0.1 0.8 Fusion protein 22 67.8 23.7 6.9 10.8 Fusion protein 23 91.3 0.9 −0.3 3.2 Fusion protein 24 82.8 16.6 16 9.1 Fusion protein 25 98.5 0.6 0.1 3.3 Fusion protein 26 67.2 32 12.3 14.2 Fusion protein 27 97.6 0.8 0.1 3.7 Fusion protein 28 23 77 50.2 51.1 Fusion protein 29 97.5 0.1 0.1 2.5

In summary, the superior stability of the fusion protein containing the preferred truncated TGFβR2 extracellular domain was further validated by the fusion proteins with multiple kinds of solid tumor antigens as the targeting portion.

Example 8: In Vitro Biological Property of X/TGFβR2 Fusion Proteins with Targeting Solid Tumors Antigen as Targeting Portion and Truncated TGFβR2 (Fusion Protein 6)

8.1 X/TGFβR2 Fusion Protein Binding TGF-β1 Assay

The binding ability of the X/TGFβR2 fusion protein to TGF-β1 was measured by ELISA according to Example 3.4. As shown in FIG. 33 , the ability of the X/TGFβR2 fusion protein with the preferred truncated form of TGFβR2 to bind TGF-β1 is slightly lower than that of the X/TGFβR2 fusion protein with the full-length form of TGFβR2.

8.2 X/TGFβR2 Fusion Protein Neutralizing TGF-β Assay

The ability of the X/TGFβR2 fusion protein to neutralize TGF-β1 and TGF-β3 was tested with reference to Example 3.5. As shown in FIG. 34 , the ability of the X/TGFβR2 fusion protein with the preferred truncated form of TGFβR2 to neutralize both TGF-β1 and TGF-β3 was superior to that of the X/TGFβR2 fusion protein with the full-length form of TGFβR2.

8.3 Binding Assay of X/TGFβR2 Fusion Protein for the Target of X Portion

The antigens ERBB2-his, VEGF165, VEGFR2-His, CTLA4-his, and EGFR-His, which are X portion target, at final concentrations of 10 ng/mL, 5 ng/mL, 80 ng/mL, 80 ng/mL, and 40 ng/mL, respectively, were coated on 96-well plates at 100 L/well. The plates were coated overnight at 4° C. The plates were washed the next day, blocked at room temperature for 1 h, and incubated with 100 μL of X/TGFβR2 fusion protein at a final concentration of 13.89 nM for 1 h. The plates were washed to remove unbound antibodies, incubated with secondary antibody Goat anti-hIgG Fc/HRP, and repeated the wash. Finally, the substrate chromogenic solution was added for color development, and the OD₄₅₀ was read by the a microplate reader after termination. As shown in FIG. 35 , the X/TGFβR2 fusion protein with the elected truncated TGFβR2 has a similar ability to bind the corresponding antigen on the X side as the X/TGFβR2 fusion protein containing the full-length form of TGFβR2.

SEQUENCE LISTING

Number Name Sequence SEQ The HNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNPEGKYSFGAT ID. amino SLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLL NO. sequence IQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISG 1 Human NKNLCYAMRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFEDHFLSLQ EGFRprotein RMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENLQIIRGNM (UniProtKB- YYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQWRDIVSS P35968) DFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGRCRGKSPS extracellular DCCCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGIGIGEFK domain DNTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVS acid of RGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGP Met1-Ser645 HCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPS SEQ The TCTAGTGGTGGCGGTGGTTCGGGCGGTGGTGGAGGTGGTAGTTCTAGATCTTC ID. nucleotide C NO. sequence 2 encoding the linker used in the construction of the phage antibody library for the linkage of the murine antibody scFv SEQ The The nucleotide sequence encoding EGFR-mhPA8 light ID. nucleotide chain variable region (SEQ ID NO. sequence NO: 5): 3 encoding GATATCCAGATGACCCAGTCTCCAGCCTCCCTGGCTGCATCTGTGGGAGAAAC murine TGTCACCATCACATGTCGAGCAAGTGAGAACATTTACTACAGTTTAGCTTGGTATCAG antibody scFv CAGAAGGAAGGGAAATCTCCTCAGCTCCTGATCTATATTACAAACGGCTTGGCAGAT used in the GGTGTCCCATCGAGGTTCAGTGGCAGTGGATCTGGGACACAGTATTCTATGAAGATC construction of GACAGCATGCAGCCTGAAGATACCGCAACTTATTTCTGTAAACAGTCTTATGACGTTC antibody CGCTCACGTTCGGTGCTGGGACCAAGCTGGAGATGAAA EGFR-mhPA8 Linker (SEQ ID NO: 2): TCTAGTGGTGGCGGTGGTTCGGGCGGTGGTGGAGGTGGTAGTTCTAGATCTTC C The nucleotide sequence encoding EGFR-mhPA8 light heavy variable region (SEQ ID NO: 4): CAGGTGCAGCTGCAGCAATCTGGACCTGATTTGGTGAAGCCTGGGGCTTCAG TGAGGATATCTTGCAAGGTTTCTGGCTACACCTTCACAACCTACTATACACACTGGGT GAAGCAGAGGCCTGGACGGGGACTTGAATGGATTGGATGGATTTATCCTGGAGATGT TAATACGAAGTACAATGAGAAATTCAAGGGCAAGGCCACACTGACTGCAGACAAAA CCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCT ATTTCTGTGCAAGAGAAGACCCCGGTAGTAACTACTTTGACTACTGGGGCCAAGGCA CCACTCTCACAGTCTCCTCA SEQ The CAGGTGCAGCTGCAGCAATCTGGACCTGATTTGGTGAAGCCTGGGGCTTCAG ID. nucleotide TGAGGATATCTTGCAAGGTTTCTGGCTACACCTTCACAACCTACTATACACACTGGGT NO: sequence GAAGCAGAGGCCTGGACGGGGACTTGAATGGATTGGATGGATTTATCCTGGAGATGT 4 encoding TAATACGAAGTACAATGAGAAATTCAAGGGCAAGGCCACACTGACTGCAGACAAAA heavy CCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCT variable ATTTCTGTGCAAGAGAAGACCCCGGTAGTAACTACTTTGACTACTGGGGCCAAGGCA region CCACTCTCACAGTCTCCTCA of murine antibody EGFR-mhPA8 the chain SEQ The GATATCCAGATGACCCAGTCTCCAGCCTCCCTGGCTGCATCTGTGGGAGAAAC ID. nucleotide TGTCACCATCACATGTCGAGCAAGTGAGAACATTTACTACAGTTTAGCTTGGTATCAG NO: sequence CAGAAGGAAGGGAAATCTCCTCAGCTCCTGATCTATATTACAAACGGCTTGGCAGAT 5 encoding GGTGTCCCATCGAGGTTCAGTGGCAGTGGATCTGGGACACAGTATTCTATGAAGATC light GACAGCATGCAGCCTGAAGATACCGCAACTTATTTCTGTAAACAGTCTTATGACGTTC variable CGCTCACGTTCGGTGCTGGGACCAAGCTGGAGATGAAA region of murine antibody EGFR-mhPA8 SEQ Nucleotide GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA ID. sequence CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG NO: of human GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC 6 IgG1 TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG heavy CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA chain AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG constant TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC region AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTC'AACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGAGCAAGCACCAAGGGCCCA TCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ Nucleotide CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTT ID. sequence GAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC NO: of human AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC 7 kappa ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAG light CAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCC chain TGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG constant region SEQ The QVQLQQSGPDLVKPGASVRISCKVSGYTFTTYYTHWVKQRPGRGLEWIGWIYP ID. amino acid GDVNTKYNEKFKGKATLTADKTSSTAYMQLSSLTSEDSAVYFCAREDPGSNYFDYWGQ NO: sequence GTTLTVSS 8 of the heavy chain variable region of murine antibody EGFR- mhPA8 SEQ The DIQMTQSPASLAASVGETVTITCRASENIYYSLAWYQQKEGKSPQLLIYITNGLA ID. amino acid DGVPSRFSGSGSGTQYSMKIDSMQPEDTATYFCKQSYDVPLTFGAGTKLEMK NO: sequence 9 of the light chain variable region of murine antibody EGFR- mhPA8 SEQ The RASENIYYSLA ID. amino acid NO: sequence 10 of murine antibody EGFR- mhPA8 light chain CDR1 SEQ The ITNGLAD ID. amino acid NO: sequence of 11 murine antibody EGFR-mhPA8 light chain CDR2 SEQ The KQSYDVPLT ID. amino acid NO: sequence 12 of murine antibody EGFR- mhPA8 light chain CDR3 SEQ The GYTFTTYYTH ID. amino acid NO: sequence 13 of murine antibody EGFR- mhPA8 heavy chain CDR1 SEQ The WIYPGDVNTKYNEKFKG ID. amino acid NO: sequence 14 of murine antibody EGFR- mhPA8 heavy chain CDR2 SEQ The AREDPGSNYFDY ID. amino acid NO: sequence 15 of murine antibody EGFR- mhPA8 heavy chain CDR3 SEQ The RASENIYYSLA ID. amino acid NO: sequence of 16 humanized antibody EGFR-HPA8 light chain CDR1 SEQ The ITDGLAD ID. amino acid NO: sequence of 17 humanized antibody EGFR-HPA8 light chain CDR2 SEQ The KQSYDVPLT ID. amino acid NO: sequence 18 humanized antibody EGFR- HPA8 light chain CDR3 SEQ The GYTFTTYYTH ID. amino acid NO: sequence 19 of humanized antibody EGFR- HPA8 heavy chain CDR1 SEQ The WIYPGDVNTKYNEKFKG ID. amino acid NO: sequence 20 of humanized antibody EGFR- HPA8 heavy chain CDR2 SEQ The AREDPGSNYFDY ID. amino acid NO: sequence of 21 humanized antibody EGFR-HPA8 heavy chain CDR3 SEQ The The amino acid sequence of the heavy ID. amino acid chain variable region (SEQ ID NO: 28): NO: sequence of EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP 22 humanized GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ antibody GTLVTVSS EGFR-HPA8 The amino acid sequence of the heavy heavy chain chain constant region (SEQ ID NO: 30): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ The The amino acid sequence of the light ID. amino acid chain variable region (SEQ ID NO: 29): NO: sequence of DIQMTQSPSSLSASVGDRVTITCRASENIYYSLAWYQQKPGKAPQLLIYITDGLA 23 humanized DGVPSRFSGSGSGTDYTLKIDSLQPEDFATYYCKQSYDVPLTFGGGTKVEIK antibody The amino acid sequence of the light EGFR-HPA8 chain constant regio (SEQ ID NO: 31): light chain RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ The The amino acid sequence of the heavy ID. amino acid chain signal peptide (SEQ ID NO: 26): NO: sequence of MGWSLILLFLVAVATRVLS 24 humanized The amino acid sequence of the heavy antibody chain variable region (SEQ ID NO: 28): EGFR-HPA8 EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP heavy chain GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ containing the GTLVTVSS signal peptide The amino acid sequence of the heavy chain constant region (SEQ ID NO: 30): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ The The amino acid sequence of the light ID. amino acid chain signal peptide (SEQ ID NO: 27): NO: sequence of MGWSCIILFLVATATGVHS 25 humanized The amino acid sequence of the light antibody chain variable region (SEQ ID NO: 29): EGFR-HPA8 DIQMTQSPSSLSASVGDRVTITCRASENIYYSLAWYQQKPGKAPQLLIYITDGLA light chain DGVPSRFSGSGSGTDYTLKIDSLQPEDFATYYCKQSYDVPLTFGGGTKVEIK containing a The amino acid sequence of the light signal peptide chain constant region (SEQ ID NO: 31): RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ The MGWSLILLFLVAVATRVLS ID. amino acid NO: sequence of 26 humanized antibodyEGF R-HPA8 heavy chain signal peptide SEQ The MGWSCIILFLVATATGVHS ID. amino acid NO: sequence 27 humanized antibody EGFR- HPA8light chain signal peptide SEQ The EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP ID. amino acid GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ NO: sequence GTLVTVSS 28 of the humanized antibody EGFR- HPA8 heavy chain variable region SEQ The DIQMTQSPSSLSASVGDRVTITCRASENIYYSLAWYQQKPGKAPQLLIYITDGLA ID. amino acid DGVPSRFSGSGSGTDYTLKIDSLQPEDFATYYCKQSYDVPLTFGGGTKVEIK NO: sequence 29 of the humanized antibody EGFR- HPA8 light chain variable region SEQ The ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA ID. amino acid VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP NO: sequence ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP 30 of the REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT humanized LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT antibody VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK EGFR-HPA8 heavy chain constant region SEQ The RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE ID. Amino acid SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC NO: sequence of 31 the humanized antibody EGFR- HPA8light chain constant region SEQ The The nucleotide sequence encoding the heavy ID. nucleotide chain signal peptide (SEQ ID NO: 34): NO: sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT 32 encoding GAGT humanized The nucleotide sequence encoding the heavy antibody chain variable region (SEQ ID NO: 36): EGFR-HPA8 GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA heavy chain GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG containing TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG signal peptide AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC aTGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 38): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACSTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ The The nucleotide sequence encoding the light ID. nucleotide chain signal peptide (SEQ ID NO: 35): NO: sequence ATGGGCTGGTCCTGTATCATCCTGTTCCTGGTGGCTACAGCCACAGGAGTGCA 33 encoding TAGT humanized The nucleotide sequence encoding the light antibody chain variable region (SEQ ID NO: 37): EGFR-HPA8 GACATCCAGATGACCCAGAGCCCATCCTCCCTGTCTGCCTCTGTGGGAGACA light GGGTGACCATCACTTGTAGGGCATCTGAGAACATCTACTACTCCCTGGCTTGGTATCA chain ACAGAAGCCTGGCAAGGCTCCACAGCTGCTGATTTACATCACCGACGGACTGGCTGA containing a TGGAGTGCCAAGCAGGTTCTCTGGCTCTGGCTCTGGCACAGACTACACCCTGAAGAT signal peptide CGACTCCCTCCAACCTGAGGACTTTGCCACCTACTACTGTAAGCAGTCCTATGATGTG CCACTGACCTTTGGAGGAGGCACCAAGGTGGAGATTAAG The nucleotide sequence encoding the light chain constant region (SEQ ID NO: 39): CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTT GAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAG CAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCC TGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTGA SEQ The ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT ID. nucleotide GAGT NO: sequence 34 encoding humanized antibody EGFR- HPA8 heavy chain signal peptide SEQ The ATGGGCTGGTCCTGTATCATCCTGTTCCTGGTGGCTACAGCCACAGGAGTGCA ID. nucleotide TAGT NO: sequence 35 encoding humanized antibody EGFR- HPA8 light chain signal peptide SEQ The GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA ID. nucleotide GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG NO: sequence TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG 36 encoding TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC humanized AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC antibody TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC EGFR-HPA8 ACCCTGGTGACAGTGTCCAGC heavy chain variable region SEQ The GACATCCAGATGACCCAGAGCCCATCCTCCCTGTCTGCCTCTGTGGGAGACA ID. nucleotide GGGTGACCATCACTTGTAGGGCATCTGAGAACATCTACTACTCCCTGGCTTGGTATCA NO: sequence ACAGAAGCCTGGCAAGGCTCCACAGCTGCTGATTTACATCACCGACGGACTGGCTGA 37 encoding TGGAGTGCCAAGCAGGTTCTCTGGCTCTGGCTCTGGCACAGACTACACCCTGAAGAT humanized CGACTCCCTCCAACCTGAGGACTTTGCCACCTACTACTGTAAGCAGTCCTATGATGTG antibody CCACTGACCTTTGGAGGAGGCACCAAGGTGGAGATTAAG EGFR- HPA8 light chain variable region SEQ The GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA ID. nucleotide CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG NO: sequence GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC 38 encoding TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG humanized CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA antibody AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG EGFR-HPA8 TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC heavy AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG constant AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA region TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA chain GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ The CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTT ID. nucleotide GAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC NO: sequence AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC 39 encoding ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAG humanized CAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCC antibody TGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTGA EGFR-HPA8 light chain constant region SEQ The The amino acid sequence of the EGFR-mhPA8 light ID. amino acid chain variable region (SEQ ID NO: 9): NO: sequence DIQMTQSPASLAASVGETVTITCRASENIYYSLAWYQQKEGKSPQLLIYITNGLA 40 of murine DGVPSRFSGSGSGTQYSMKIDSMQPEDTATYFCKQSYDVPLTFGAGTKLEMK antibody scFv Linker (SEQ ID NO: 41): used in the SSGGGGSGGGGGGSSRSS construction of The amino acid sequence of the EGFR-mhPA8 heavy antibody chain variable region (SEQ ID EGFR-mhPA8 NO: 8): GDVNTKYNEKFKGKATLTADKTSSTAYMQLSSLTSEDSAVYFCAREDPGSNYFDYWGQ QVQLQQSGPDLVKPGASVRISCKVSGYTFTTYYTHWVKQRPGRGLEWIGWIYP GTTLTVSS SEQ The SSGGGGSGGGGGGSSRSS ID. amino acid NO: sequence of 41 the linker used in the construction of the phage antibody library for the linkage of the murine antibody scFv SEQ The The nucleotide sequence encoding the heavy ID. nucleotide chain signal peptide (SEQ ID NO: 34): NO: sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT 42 encoding GAGT human-mouse The nucleotide sequence encoding the heavy chimeric chain variable region (SEQ ID NO: 4): antibody CAGGTGCAGCTGCAGCAATCTGGACCTGATTTGGTGAAGCCTGGGGCTTCAG EGFR-mhPA8 TGAGGATATCTTGCAAGGTTTCTGGCTACACCTTCACAACCTACTATACACACTGGGT heavy chain GAAGCAGAGGCCTGGACGGGGACTTGAATGGATTGGATGGATTTATCCTGGAGATGT containing the TAATACGAAGTACAATGAGAAATTCAAGGGCAAGGCCACACTGACTGCAGACAAAA signal peptide CCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCT ATTTCTGTGCAAGAGAAGACCCCGGTAGTAACTACTTTGACTACTGGGGCCAAGGCA CCACTCTCACAGTCTCCTCA The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 6): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ The The nucleotide sequence encoding the light ID. nucleotide chain signal peptide (SEQ ID NO: 35): NO: sequence ATGGGCTGGTCCTGTATCATCCTGTTCCTGGTGGCTACAGCCACAGGAGTGCA 43 encoding TAGT human-mouse The nucleotide sequence encoding the light chimeric chain variable region (SEQ ID NO: 5): antibody GATATCCAGATGACCCAGTCTCCAGCCTCCCTGGCTGCATCTGTGGGAGAAAC EGFR-mhPA8 TGTCACCATCACATGTCGAGCAAGTGAGAACATTTACTACAGTTTAGCTTGGTATCAG light chain CAGAAGGAAGGGAAATCTCCTCAGCTCCTGATCTATATTACAAACGGCTTGGCAGAT containing the GGTGTCCCATCGAGGTTCAGTGGCAGTGGATCTGGGACACAGTATTCTATGAAGATC signal peptide GACAGCATGCAGCCTGAAGATACCGCAACTTATTTCTGTAAACAGTCTTATGACGTTC CGCTCACGTTCGGTGCTGGGACCAAGCTGGAGATGAAA The nucleotide sequence encoding the light chain constant region (SEQ ID NO: 7): CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTT GAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAG CAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCC TGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ The The amino acid sequence of the heavy ID. amino acid chain signal peptide (SEQ ID NO: 26): NO: sequence of MGWSLILLFLVAVATRVLS 44 human-mouse The amino acid sequence of the heavy chimeric chain variable region (SEQ ID NO: 8): antibody QVQLQQSGPDLVKPGASVRISCKVSGYTFTTYYTHWVKQRPGRGLEWIGWIYP EGFR-mhPA8 GDVNTKYNEKFKGKATLTADKTSSTAYMQLSSLTSEDSAVYFCAREDPGSNYFDYWGQ heavy chain GTTLTVSS containing the The amino acid sequence of the heavy signal peptide chain constant region (SEQ ID NO: 30): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ The The amino acid sequence of the light ID. amino acid chain signal peptide (SEQ ID NO: 27): NO: sequence of MGWSCIILFLVATATGVHS 45 human-mouse The amino acid sequence of the light chimeric chain variable region (SEQ ID NO: 9): antibody DIQMTQSPASLAASVGETVTITCRASENIYYSLAWYQQKEGKSPQLLIYITNGLA EGFR-mhPA8 DGVPSRFSGSGSGTQYSMKIDSMQPEDTATYFCKQSYDVPLTFGAGTKLEMK light chain The amino acid sequence of the light containing the chain constant region (SEQ ID NO: 31): signal peptide RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID The ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA NO: amino acid VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP 46 sequence of ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP humanized REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT antibody LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT EGFR-HPA8 VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG heavy chain constant region with C- terminal lysine deletion SEQ ID The IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE NO: amino acid KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS 47 sequence CSSDECNDNIIFSEEYNTSNPD of the full-length TGFβR2 extracellular domain SEQ ID The IPPHVNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRK NO: amino acid NDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSE 48 sequence of EYNTSNPD truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-16) SEQ ID The IPPHVGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKND NO: amino acid ENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEY 49 sequence of NTSNPD truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-19) SEQ ID The IPPHVVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDEN NO: 50 amino acid ITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNT sequence of SNPD truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-21) SEQ ID The IPPHVQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLE NO: 51 amino acid TVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-25) SEQ ID The IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET NO: 52 amino acid VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-26) acid SEQ ID The IPPHVCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETV NO: 53 amino acid CHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (6-27) SEQ ID The IPPHVQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLE NO: 54 amino acid TVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (7-26) SEQ ID The IPPHLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVC NO: 55 amino acid HDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (5-26) SEQ ID The IPPLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH NO: 56 amino acid DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (4-26) acid SEQ ID The IPLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH NO: 57 amino DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (3-26) SEQ ID The ILCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHD NO: 58 amino acid PKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (2-26) SEQ ID The PPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETV NO: 59 amino acid CHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- erminal amino acids at position (1, 6-26) SEQ ID The PHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVC NO: 60 amino acid HDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1- 2, 6-26) SEQ ID The HVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH NO: 61 amino acid DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1- 3, 6-26) SEQ ID The VLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHD NO: 62 amino acid PKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1- 4, 6-26) SEQ ID The GAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITL NO: 63 amino acid ETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEE sequence of YNTSNPD truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1-19) SEQ ID The VKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET NO: 64 amino acid VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIF sequence of SEEYNTSNPD truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1-21) SEQ ID The LCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDP NO: 65 amino acid KLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD sequence of truncated TGFβR2 extracellular domain with a deletion of N- terminal amino acids at position (1-26) SEQ ID The GGGGSGGGGSGGGGSGGGGS NO: 66 amino acid sequence of Linker linking the heavy chain C- terminal and TGFβR2 extracellular domain in fusion protein SEQ ID The GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGC NO: 67 nucleotide ACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGG sequence CCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAA encoding CCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGAC humanized CAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGT antibody CCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACC EGFR-HPA8 GTGCCCTCCAGCAGCTTGGGCACCCAGACCTACAT heavy CTGCAACGTGAATCACAAGCCCAGCAACACCAAGG constant TGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAA region ACTCACACATGCCCACCGTGCCCAGCACCTGAACT with CCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAA terminal AACCCAAGGACACCCTCATGATCTCCCGGACCCCT lysine GAGGTCACgTGCGTGGTGGTGGACGTGAGCCACGA deletion AGACCCcGAGGTCAAGTTCAACTGGTACGTGGACG chain GCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGG GAGGAGCAGTACAACAGCACGTACCGTGTGGTCAG CGTCCTCACCGTCCTGCACCAGGACTGGCTGAATG GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCC CTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGC CAAAGGGCAGCCCCGAGAACCACAGGTGTACACCC TGCCCCCATCCCGGGATGAGCTGACCAAGAACCAG GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCC CAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC AGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAA GCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGA ACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG CACAACCACTACACGCAGAAGAGCCTCTCCCTGTC TCCGGGT SEQ ID The ATCCCACCGCACGTTCAGAAGTCGGTTAATAACGA NO: 68 nucleotide CATGATAGTCACTGACAACAACGGTGCAGTCAAGT sequence TTCCACAACTGTGTAAATTTTGTGATGTGAGATTT encoding TCCACCTGTGACAACCAGAAATCCTGCATGAGCAA full- CTGCAGCATCACCTCCATCTGTGAGAAGCCACAGG length the AAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAG extracellular AACATAACACTAGAGACAGTTTGCCATGACCCCAA domain of GCTCCCCTACCATGACTTTATTCTGGAAGATGCTG TGFβR2 CTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAG CCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTC TGATGAGTGCAATGACAACATCATCTTCTCAGAAG AATATAACACCAGCAATCCTGACTAAA SEQ ID The ATCCCACCGCACGTTAACAACGGTGCAGTCAAGTT NO: 69 nucleotide TCCACAACTGTGTAAATTTTGTGATGTGAGATTTT sequence CCACCTGTGACAACCAGAAATCCTGCATGAGCAAC encoding TGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA truncated AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGA TGFβR2 ACATAACACTAGAGACAGTTTGCCATGACCCCAAG extracellular CTCCCCTACCATGACTTTATTCTGGAAGATGCTGC domain with a TTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGC deletion of N- CTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT terminal amino GATGAGTGCAATGACAACATCATCTTCTCAGAAGA acids at ATATAACACCAGCAATCCTGACTAAA position (6-16) SEQ ID The ATCCCACCGCACGTTGGTGCAGTCAAGTTTCCACA NO: 70 nucleotide ACTGTGTAAATTTTGTGATGTGAGATTTTCCACCT sequence GTGACAACCAGAAATCCTGCATGAGCAACTGCAGC encoding ATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG truncated TGTGGCTGTATGGAGAAAGAATGACGAGAACATAA TGFβR2 CACTAGAGACAGTTTGCCATGACCCCAAGCTCCCC extracellular TACCATGACTTTATTCTGGAAGATGCTGCTTCTCC domain with a AAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTG deletion of N- AGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG terminal amino TGCAATGACAACATCATCTTCTCAGAAGAATATAA acids at CACCAGCAATCCTGACTAAA position (6-19) SEQ ID The ATCCCACCGCACGTTGTCAAGTTTCCACAACTGTG NO: 71 nucleotide TAAATTTTGTGATGTGAGATTTTCCACCTGTGACA sequence ACCAGAAATCCTGCATGAGCAACTGCAGCATCACC encoding TCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGC truncated TGTATGGAGAAAGAATGACGAGAACATAACACTAG TGFβR2 AGACAGTTTGCCATGACCCCAAGCTCCCCTACCAT extracellular GACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTG domain with a CATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTT deletion of N- TCTTCATGTGTTCCTGTAGCTCTGATGAGTGCAAT terminal amino GACAACATCATCTTCTCAGAAGAATATAACACCAG acids at CAATCCTGACTAAA position (6-21) SEQ ID The ATCCCACCGCACGTTCAACTGTGTAAATTTTGTGA NO: 72 nucleotide TGTGAGATTTTCCACCTGTGACAACCAGAAATCCT sequence GCATGAGCAACTGCAGCATCACCTCCATCTGTGAG encoding AAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAA runcated GAATGACGAGAACATAACACTAGAGACAGTTTGCC TGFβR2 ATGACCCCAAGCTCCCCTACCATGACTTTATTCTG extracellular GAAGATGCTGCTTCT domain with a CCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGG deletion of N- TGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATG terminal amino AGTGCAATGACAACATCATCTTCTCAGAAGAATAT acids at AACACCAGCAATCCTGACTAAA position (6-25) SEQ ID The ATCCCACCGCACGTTCTGTGTAAATTTTGTGATGT NO: 73 nucleotide GAGATTTTCCACCTGTGACAACCAGAAATCCTGCA sequence TGAGCAACTGCAGCATCACCTCCATCTGTGAGAAG encoding CCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAA truncated TGACGAGAACATAACACTAGAGACAGTTTGCCATG TGFβR2 ACCCCAAGCTCCCCTACCATGACTTTATTCTGGAA extracellular GATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAA domain with a AAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCT deletion of N- GTAGCTCTGATGAGTGCAATGACAACATCATCTTC terminal amino TCAGAAGAATATAACACCAGCAATCCTGACTAAA acids at position (6-26) SEQ ID The ATCCCACCGCACGTTTGTAAATTTTGTGATGTGAG NO: 74 nucleotide ATTTTCCACCTGTGACAACCAGAAATCCTGCATGA sequence GCAACTGCAGCATCACCTCCATCTGTGAGAAGCCA encoding CAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGA truncated CGAGAACATAACACTAGAGACAGTTTGCCATGACC TGFβR2 CCAAGCTCCCCTACCATGACTTTATTCTGGAAGAT extracellular GCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAA domain with a AAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTA deletion of N- GCTCTGATGAGTGCAATGACAACATCATCTTCTCA terminal amino GAAGAATATAACACCAGCAATCCTGACTAAA acids at position (6-27) SEQ ID The ATCCCACCGCACGTTCAGCTGTGTAAATTTTGTGA NO: 75 nucleotide TGTGAGATTTTCCACCTGTGACAACCAGAAATCCT sequence GCATGAGCAACTGCAGCATCACCTCCATCTGTGAG encoding AAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAA truncated GAATGACGAGAACATAACACTAGAGACAGTTTGCC TGFβR2 ATGACCCCAAGCTCCCCTACCATGACTTTATTCTG extracellular GAAGATGCTGCTTCTCCAAAGTGCATTATGAAGGA domain with a AAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTT deletion of N- CCTGTAGCTCTGATGAGTGCAATGACAACATCATC terminal amino TTCTCAGAAGAATATAACACCAGCAATCCTGACTA acids at AA position (7-26) SEQ ID The ATCCCACCGCACCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA NO: 76 nucleotide CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA sequence AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG encoding CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG truncated TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TGFØR2 TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG extracellular ACTAAA domain with a deletion of N- terminal amino acids at position (5-26) SEQ ID The ATCCCACCGCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA NO: 77 nucleotide GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT sequence CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA encoding TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC truncated ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT TGFβR2 GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT extracellular AAA domain with a deletion of N- terminal amino acids at position (4-26) SEQ ID The ATCCCACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA NO: 78 nucleotide ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG sequence TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA encoding CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT truncated ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT TGFβR2 GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA extracellular domain with a deletion of N- terminal amino acids at position (3-26) SEQ ID The ATCCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC NO: 79 nucleotide CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT sequence GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC encoding CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG truncated AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGFβR2 TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA extracellular domain with a deletion of N- erminal amino acids at position (2-26) SEQ ID The CCACCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA NO: 80 nucleotide CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA sequence AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG encoding CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG truncated TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TGFβR2 TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG extracellular ACTAAA domain with a deletion of N- terminal amino acids at position (1, 6- 26) SEQ ID The CCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA NO: 81 nucleotide GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT sequence CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA encoding TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC truncated ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT TGFβR2 GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT extracellular AAA domain with a deletion of N- terminal amino acids at position (1- 2, 6-26) SEQ ID The CACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA NO: 82 nucleotide ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG sequence TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA encoding CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT truncated ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT TGFβR2 GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA extracellular domain with a deletion of N- terminal amino acids at position (1- 3, 6-26) SEQ ID The GTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC NO: 83 nucleotide CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT sequence GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC encoding CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG truncated AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGFβR2 TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA extracellular domain with a deletion of N- terminal amino acids at position (1- 4, 6-26) SEQ ID The GGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCAC NO: 84 nucleotide CTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAA sequence GCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAG encoding AGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGC truncated TTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTG TGFβR2 TTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACC extracellular AGCAATCCTGACTAAA domain with a deletion of N- terminal amino lacids at position (1-19) SEQ ID The GTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGA NO: 85 nucleotide CAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACA sequence GGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAG encoding TTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCC truncated AAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTG TGFβR2 TAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAAT extracellular CCTGACTAAA domain with a deletion of N- terminal amino acids at position (1-21) SEQ ID The CTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATCCTG NO: 86 nucleotide CATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGC sequence TGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCCCAA encoding GCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAG truncated GAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGC TGFβR2 AATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA extracellular domain with a deletion of N- terminal amino acids at position (1-26) SEQ ID The GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT NO: 87 nucleotide GGTGGTTCT sequence encoding Linker linking the heavy chain C- terminal and TGFβR2 extracellular domain in the fusion protein SEQ ID The The amino acid sequence of the heavy chain signal NO: 88 amino acid peptide (SEQ ID NO: 26): sequence MGWSLILLFLVAVATRVLS of the The amino acid sequence of the heavy chain signal variable region (SEQ ID NO: 28): peptide EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP bearing GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ heavy GTLVTVSS chain of The amino acid sequence of the heavy chain fusion constant region (SEQ ID NO: 46): protein 1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy chain signal NO: 89 amino acid peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain variable bearing heavy region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 2 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 48): IPPHVNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRK NDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSE EYNTSNPD SEQ ID The The amino acid sequence of the heavy chain signal NO: 90 amino acid peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain bearing heavy variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 3 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 49): IPPHVGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKND ENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEY NTSNPD SEQ ID The The amino acid sequence of the heavy chain NO: 91 amino acid signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain variable bearing heavy region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein4 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 50): IPPHVVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDEN ITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNT SNPD SEQ ID The The amino acid sequence of the heavy chain signal NO: 92 amino acid peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain variable bearing heavy region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein5 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 51): IPPHVQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLE TVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy chain signal NO: 93 amino acid peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain variable bearing heavy region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein6 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy chain signal NO: 94 amino acid peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy chain variable bearing heavy region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein7 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 53): IPPHVCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETV CHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID amino The amino acid sequence of the heavy chain signal NO: 95 The peptide (SEQ ID NO: 26): acid MGWSLILLFLVAVATRVLS sequence The amino acid sequence of the heavy of the chain variable region (SEQ ID NO: 28): signal EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP peptide GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ bearing GTLVTVSS heavy The amino acid sequence of the heavy chain of chain constant region (SEQ ID NO: 46): fusion ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA protein8 VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 54): IPPHVQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLE TVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 96 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 9 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 55): IPPHLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVC HDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 97 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 10 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 56): IPPLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 98 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 11 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 57): IPLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 99 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 12 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 58): ILCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHD PKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 100 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 13 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 59): PPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETV CHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 101 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 14 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 60): PHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVC HDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 102 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 15 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 61): HVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCH DPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 103 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 16 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 62): VLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHD PKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 104 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 17 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 63): GAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITL ETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNP D SEQ ID The The amino acid sequence of the heavy NO: 105 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 18 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 64): VKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 106 amino acid chain signal peptide (SEQ ID NO: 26): sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region (SEQ ID NO: 28): chain of fusion EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP protein 19 GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full-length TGFβR2 extracellular domain (SEQ ID NO: 65): LCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDP KLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The nucleotide sequence encoding the heavy NO: 107 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 1 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCCGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full-length extracellular domain of TGFβR2 (SEQ ID NO: 68): ATCCCACCGCACGTTCAGAAGTCGGTTAATAACGACATGATAGTCACTGACAA CAACGGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCACC TGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAG CCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGA GACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCT TCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGT TCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCA GCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 108 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding GAGT fusion protein The nucleotide sequence encoding the heavy the signal chain variable region (SEQ ID NO: 36): peptide GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA bearing heavy GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG chain of fusion TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG protein 2 TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 69): ATCCCACCGCACGTTAACAACGGTGCAGTCAAGTTTCCACAACTGTGTAAATT TTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAG CATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAA TGACGAGAACATAACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGA CTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCC TGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATC TTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 109 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 3 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full-length extracellular domain of TGFβR2 (SEQ ID NO: 70): ATCCCACCGCACGTTGGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGA TGTGAGATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCAC CTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGA GAACATAACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATT CTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGA GACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAG AAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 110 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 4 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 71): ATCCCACCGCACGTTGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAG ATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATC TGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACAT AACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAA GATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTC TTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAAT ATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 111 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein5 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 72): ATCCCACCGCACGTTCAACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGT GACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCA CAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGAC AGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCT CCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCC TGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCA ATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 112 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein6 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 73): ATCCCACCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGAC AACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAG GAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTT TGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAA AGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTA GCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCC TGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 113 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein7 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 74): ATCCCACCGCACGTTTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 114 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein8 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domainof TGFβR2 (SEQ ID NO: 75): ATCCCACCGCACGTTCAGCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGT GACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCA CAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGAC AGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCT CCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCC TGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCA ATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 115 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 9 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 76): ATCCCACCGCACCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 116 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 10 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 77: ATCCCACCGCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT AAA SEQ ID The The nucleotide sequence encoding the heavy NO: 117 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 11 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 78): ATCCCACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 118 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 12 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 79): ATCCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 119 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 13 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 80): CCACCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 120 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 14 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 81): CCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT AAA SEQ ID The The nucleotide sequence encoding the heavy NO: 121 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 15 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 82): CACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 122 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 16 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 83): GTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 123 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding, the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 17 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 84): GGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCAC CTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAA GCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAG AGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGC TTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTG TTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACC AGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 124 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 18 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 85): GTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGA CAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACA GGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAG TTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCC AAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTG TAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAAT CCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 125 nucleotide chain signal peptide (SEQ ID NO: 34): sequence ATGGGCTGGTCCCTGATTCTGCTGTTCCTGGTGGCTGTGGCTACCAGGGTGCT encoding the GAGT signal peptide The nucleotide sequence encoding the heavy bearing heavy chain variable region (SEQ ID NO: 36): chain of fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 19 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 86): CTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATCCTG CATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGC TGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCCCAA GCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAG GAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGC AATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The amino acid sequence of the light chain signal peptide: NO: 126 amino acid MGWSLILLFLVAVATRVLS sequence of the The amino acid sequence of the light chain variable region: signal peptide DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLY bearing light SGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK chain of fusion The amino acid sequence of the light chain constant region: protein 20, 21 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID The The amino acid sequence of the heavy NO: 127 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion EVOLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPT protein 20 NGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWG QGTLVTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 128 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPT protein 21 NGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWG QGTLVTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the light chain signal peptide: NO: 129 amino acid MGWSLILLFLVAVATRVLS sequence of the The amino acid sequence of the light chain variable region: signal peptide DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHS bearing light GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIK chain of fusion The amino acid sequence of the light chain constant region: protein 22, 23 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID The amino acid The amino acid sequence of the heavy NO: 130 sequence of the chain signal peptide: signal peptide MGWSLILLFLVAVATRVLS bearing heavy The amino acid sequence of the heavy chain of chain variable region: fusion DVWGQGTLVTVSS EVOLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEW protein 22 VGWIN TYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSS HWYF The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 131 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion EVOLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEWVGWIN protein 23 TYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSSHWYF DVWGQGTLVTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD The amino acid sequence of the light chain signal peptide: MGWSLILLFLVAVATRVLS The amino acid sequence of the light chain variable region: TGVPSRFSGSGSGTYFTLTISSLQAEDFAVYFCQQAKAFPPTFGGGTKVDIK DIQMTQSPSSVSASIGDRVTITCRASQGIDNWLGWYQQKPGKAPKLLIYDASNLD The amino acid sequence of the light chain constant region: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SEQ ID The SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC NO: 132 amino acid The amino acid sequence of the heavy sequence of the chain signal peptide: signal peptide MGWSLILLFLVAVATRVLS bearing The amino acid sequence of the heavy light chain of chain variable region: fusion EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSS protein 24, 25 SSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVTDAFDIWGQGTMV TVSS SEQ ID The The amino acid sequence of the heavy NO: 133 amino acid chain constant region sequence of the ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA signal peptide VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP bearing heavy ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP chain of fusion REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT protein 24 LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 134 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSS protein 25 SSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVTDAFDIWGQGTMV TVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the light chain signal peptide: NO: 135 amino acid MGWSLILLFLVAVATRVLS sequence of the The amino acid sequence of the light chain variable region: signal peptide EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFSRA bearing light TGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK chain of fusion The amino acid sequence of the light chain constant region: protein 26, 27 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID The The amino acid sequence of the heavy NO: 136 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVTFISY protein 26 DGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQ GTLVTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 137 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion DGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQ protein 27 GTLVTVSS QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEW VTFISY The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the light chain signal peptide: NO: 138 amino acid MGWSLILLFLVAVATRVLS sequence of the The amino acid sequence of the light chain variable region: signal peptide DIQMTQSPAALSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNL bearing light ETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLPLAFGGGTKVEIK chain of fusion The amino acid sequence of the light chain constant region: protein 28, 29 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID The The amino acid sequence of the heavy NO: 139 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGHIY protein 28 YSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTGAFDIWGQGTM VTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 47): IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCS CSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 140 amino acid chain signal peptide: sequence of the MGWSLILLFLVAVATRVLS signal peptide The amino acid sequence of the heavy bearing heavy chain variable region: chain of fusion QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGHIY protein 29 YSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTGAFDIWGQGTM VTVSS The amino acid sequence of the heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLET VCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The The amino acid sequence of the heavy NO: 141 amino acid chain variable region (SEQ ID NO: 28): sequence of EVQLVQSGAEVKKPGATVKISCKVSGYTFTTYYTHWVKQAPGKGLEWIGWIYP fusion protein GDVNTKYNEKFKGRVTLTADTSTDTAYMELSSLRSEDTAVYYCAREDPGSNYFDYWGQ 6 heavy JGTLVTVSS chain The amino acid sequence of the heavy chain constant region (SEQ ID NO: 46): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The amino acid sequence of Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 66): GGGGSGGGGSGGGGSGGGGS The amino acid sequence of the full- length TGFβR2 extracellular domain (SEQ ID NO: 52): IPPHVLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWR KNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFF MCSCSSDECNDNIIFSEEYNTSNPD SEQ ID The nucleotide ATGACAGTGCTGGCGCCAGCCTGGAGCCCAAATTCCTCCCTGTTG NO: 142 sequence CTGCTGTTGCTGCTGCTGAGTCCTTGCCTGCGGGGGACACCTGAC plasmid of TGTTACTTCAGCCACAGTCCCATCTCCTCCAACTTCAAAGTGAAG pCMV3- TTTAGAGAGTTGACTGACCACCTGCTTAAAGATTACCCAGTCACT mFlt3L GTGGCCGTCAATCTTCAGGACGAGAAGCACTGCAAGGCCTTGTGG AGCCTCTTCCTAGCCCAGCGCTGGATAGAGCAACTGAAGACTGTG GCAGGGTCTAAGATGCAAACGCTTCTGGAGGACGTCAACACCGAG ATACATTTTGTCACCTCATGTACCTTCCAGCCCCTACCAGAATGT CTGCGATTCGTCCAGACCAACATCTCCCACCTCCTGAAGGACACC TGCACACAGCTGCTTGCTCTGAAGCCCTGTATCGGGAAGGCCTGC CAGAATTTCTCTCGGTGCCTGGAGGTGCAGTGCCAGCCGGACTCC TCCACCCTGCTGCCCCCAAGGAGTCCCATAGCCCTAGAAGCCACG GAGCTCCCAGAGCCTCGGCCCAGGCAGCTGTTGCTCCTGCTGCTG CTGCTGCTGCCTCTCACACTGGTGCTGCTGGCAGCCGCCTGGGGC CTTCGCTGGCAAAGGGCAAGAAGGAGGGGGGAGCTCCACCCTGGG GTGCCCCTCCCCTCCCATCCCTAA SEQ ID The nucleotide ATGTGGCTGCAGAATTTACTTTTCCTGGGCATTGTGGTCTACAGC NO: 143 sequence of CTCTCAGCACCCACCCGCTCACCCATCACTGTCACCCGGCCTTGG plasmid AAGCATGTAGAGGCCATCAAAGAAGCCCTGAACCTCCTGGATGAC pCMV3- ATGCCTGTCACGTTGAATGAAGAGGTAGAAGTCGTCTCTAACGAG mCSF2 TTCTCCTTCAAGAAGCTAACATGTGTGCAGACCCGCCTGAAGATA TTCGAGCAGGGTCTACGGGGCAATTTCACCAAACTCAAGGGCGCC TTGAACATGACAGCCAGCTACTACCAGACATACTGCCCCCCAACT CCGGAAACGGACTGTGAAACACAAGTTACCACCTATGCGGATTTC ATAGACAGCCTTAAAACCTTTCTGACTGATATCCCCTTTGAATGC AAAAAACCAGGCCAAAAATAA SEQ ID The nucleotide ATGCGACCCTCCGGGACGGCCGGGGCAGCGCTCCTGGCGCTGCTGGCTGCGC NO: 144 sequence of TCTGCCCGGCGAGTCGGGCTCTGGAGGAAAAGAAAGTTTGCCAAGGCACGAGTAAC plasmid AAGCTCACGCAGTTGGGCACTTTTGAAGATCATTTTCTCAGCCTCCAGAGGATGTTC EGFR-TT- AATAACTGTGAGGTGGTCCTTGGGAATTTGGAAATTACCTATGTGCAGAGGAATTATG WPRE ATCTTTCCTTCTTAAAGACCATCCAGGAGGTGGCTGGTTATGTCCTCATTGCCCTCAA CACAGTGGAGCGAATTCCTTTGGAAAACCTGCAGATCATCAGAGGAAATATGTACTA CGAAAATTCCTATGCCTTAGCAGTCTTATCTAACTATGATGCAAATAAAACCGGACTG AAGGAGCTGCCCATGAGAAATTTACAGGAAATCCTGCATGGCGCCGTGCGGTTCAGC AACAACCCTGCCCTGTGCAACGTGGAGAGCATCCAGTGGCGGGACATAGTCAGCAG TGACTTTCTCAGCAACATGTCGATGGACTTCCAGAACCACCTGGGCAGCTGCCAAAA GTGTGATCCAAGCTGTCCCAATGGGAGCTGCTGGGGTGCAGGAGAGGAGAACTGCC AGAAACTGACCAAAATCATCTGTGCCCAGCAGTGCTCCGGGCGCTGCCGTGGCAAG TCCCCCAGTGACTGCTGCCACAACCAGTGTGCTGCAGGCTGCACAGGCCCCCGGGA GAGCGACTGCCTGGTCTGCCGCAAATTCCGAGACGAAGCCACGTGCAAGGACACCT GCCCCCCACTCATGCTCTACAACCCCACCACGTACCAGATGGATGTGAACCCCGAGG GCAAATACAGCTTTGGTGCCACCTGCGTGAAGAAGTGTCCCCGTAATTATGTGGTGA CAGATCACGGCTCGTGCGTCCGAGCCTGTGGGGCCGACAGCTATGAGATGGAGGAA GACGGCGTCCGCAAGTGTAAGAAGTGCGAAGGGCCTTGCCGCAAAGTGTGTAACGG AATAGGTATTGGTGAATTTAAAGACTCACTCTCCATAAATGCTACGAATATTAAACACT TCAAAAACTGCACCTCCATCAGTGGCGATCTCCACATCCTGCCGGTGGCATTTAGGG GTGACTCCTTCACACATACTCCTCCTCTGGATCCACAGGAACTGGATATTCTGAAAAC CGTAAAGGAAATCACAGGGTTTTTGCTGATTCAGGCTTGGCCTGAAAACAGGACGGA CCTCCATGCCTTTGAGAACCTAGAAATCATACGCGGCAGGACCAAGCAACATGGTCA GTTTTCTCTTGCAGTCGTCAGCCTGAACATAACATCCTTGGGATTACGCTCCCTCAAG GAGATAAGTGATGGAGATGTGATAATTTCAGGAAACAAAAATTTGTGCTATGCAAATA CAATAAACTGGAAAAAACTGTTTGGGACCTCCGGTCAGAAAACCAAAATTATAAGCA ACAGAGGTGAAAACAGCTGCAAGGCCACAGGCCAGGTCTGCCATGCCTTGTGCTCC CCCGAGGGCTGCTGGGGCCCGGAGCCCAGGGACTGCGTCTCTTGCCGGAATGTCAG CCGAGGCAGGGAATGCGTGGACAAGTGCAACCTTCTGGAGGGTGAGCCAAGGGAGT TTGTGGAGAACTCTGAGTGCATACAGTGCCACCCAGAGTGCCTGCCTCAGGCCATGA ACATCACCTGCACAGGACGGGGACCAGACAACTGTATCCAGTGTGCCCACTACATTG ACGGCCCCCACTGCGTCAAGACCTGCCCGGCAGGAGTCATGGGAGAAAACAACACC CTGGTCTGGAAGTACGCAGACGCCGGCCATGTGTGCCACCTGTGCCATCCAAACTGC ACCTACGGATGCACTGGGCCAGGTCTTGAAGGCTGTCCAACGAATGGGCCTAAGATC CCGTCCATCGCCACTGGGATGGTGGGGGCCCTCCTCTTGCTGCTGGTGGTGGCCCTG GGGATCGGCCTCTTCATGCGAAGGCGCCACATCGTTCGGAAGCGCACGCTGCGGAG GCTGCTGCAGGAGAGGGAGCTTGTGGAGCCTCTTACACCCAGTGGAGAAGCTCCCA ACCAAGCTCTCTTGAGGATCTTGAAGGAAACTGAATTCAAAAAGATCAAAGTGCTGG GCTCCGGTGCGTTCGGCACGGTGTATAAGGGACTCTGGATCCCAGAAGGTGAGAAA GTTAAAATTCCCGTCGCTATCAAGGAATTAAGAGAAGCAACATCTCCGAAAGCCAAC AAGGAAATCCTCGATGAAGCCTACGTGATGGCCAGCGTGGACAACCCCCACGTGTGC CGCCTGCTGGGCATCTGCCTCACCTCCACCGTGCAACTCATCACGCAGCTCATGCCCT TCGGCTGCCTCCTGGACTATGTCCGGGAACACAAAGACAATATTGGCTCCCAGTACCT GCTCAACTGGTGTGTGCAGATCGCAAAGGGCATGAACTACTTGGAGGACCGTCGCTT GGTGCACCGCGACCTGGCAGCCAGGAACGTACTGGTGAAAACACCGCAGCATGTCA AGATCACAGATTTTGGGCTGGCCAAACTGCTGGGTGCGGAAGAGAAAGAATACCATG CAGAAGGAGGCAAAGTGCCTATCAAGTGGATGGCATTGGAATCAATTTTACACAGAA TCTATACCCACCAGAGTGATGTCTGGAGCTACGGGGTGACCGTTTGGGAGTTGATGA CCTTTGGATCCAAGCCATATGACGGAATCCCTGCCAGCGAGATCTCCTCCATCCTGGA GAAAGGAGAACGCCTCCCTCAGCCACCCATATGTACCATCGATGTCTACATGATCATG GTCAAGTGCTGGATGATAGACGCAGATAGTCGCCCAAAGTTCCGTGAGTTGATCATC GAATTCTCCAAAATGGCCCGAGACCCCCAGCGCTACCTTGTCATTCAGGGGGATGAA AGAATGCATTTGCCAAGTCCTACAGACTCCAACTTCTACCGTGCCCTGATGGATGAAG AAGACATGGACGACGTGGTGGATGCCGACGAGTACCTCATCCCACAGCAGGGCTTCT TCAGCAGCCCCTCCACGTCACGGACTCCCCTCCTGAGCTCTCTGAGTGCAACCAGCA ACAATTCCACCGTGGCTTGCATTGATAGAAATGGGCTGCAAAGCTGTCCCATCAAGG AAGACAGCTTCTTGCAGCGATACAGCTCAGACCCCACAGGCGCCTTGACTGAGGAC AGCATAGACGACACCTTCCTCCCAGTGCCTGAATACATAAACCAGTCCGTTCCCAAA AGGCCCGCTGGCTCTGTGCAGAATCCTGTCTATCACAATCAGCCTCTGAACCCCGCG CCCAGCAGAGACCCACACTACCAGGACCCCCACAGCACTGCAGTGGGCAACCCCGA GTATCTCAACACTGTCCAGCCCACCTGTGTCAACAGCACATTCGACAGCCCTGCCCA CTGGGCCCAGAAAGGCAGCCACCAAATTAGCCTGGACAACCCTGACTACCAGCAGG ACTTCTTTCCCAAGGAAGCCAAGCCAAATGGCATCTTTAAGGGCTCCACAGCTGAAA ATGCAGAATACCTAAGGGTCGCGCCACAAAGCAGTGAATTTATTGGAGCA SEQ ID The MTVLAPAWSPNSSLLLLLLLLSPCLRGTPDCYFSHSPISSNFKVKFRELTDHLLKD NO: 145 amino acid YPVTVAVNLQDEKHCKALWSLFLAQRWIEQLKTVAGSKMQTLLEDVNTEIHFVTSC sequence TFQPLPECLRFVQTNISHLLKDTCTQLLALKPCIGKACQNFSRCLEVQCQPDSSTLL encoded by PPRSPIALEATELPEPRPRQLLLLLLLLLPLTLVLLAAAWGLRWQRARRRGELHPGVP plasmid LPSHP* pCMV3- mFlt3L SEQ ID The MWLQNLLFLGIVVYSLSAPTRSPITVTRPWKHVEAIKEALNLLDDMPVTLNEEV NO: 146 amino acid EVVSNEFSFKKLTCVQTRLKIFEQGLRGNFTKLKGALNMTASYYQTYCPPTPETDCETQ sequence VTTYADFIDSLKTFLTDIPFECKKPGQK* encoded by plasmid pCMV3- mCSF2 SEQ ID The MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFEDHFLSLQRM NO: 147 amino acid FNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENLQIIRGNMYYE sequence NSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQWRDIVSSDFLS encoded by NMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGRCRGKSPSDCC plasmid pGS6- HNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNPEGKYSFGAT EGFR-TT- CVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGIGIGEFKDSL WPRE SINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAW PENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYA NTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSR GRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHC VKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATG MVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTPSGEAPNQALLRILKET EFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSPKANKEILDEAYVMASVD NPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQYLLNWCVQIAKGMNYLE DRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHAEGGKVPIKWMALESIL KCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHLPSPTDSNFYRALMDEEDM DDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVACIDRNGLQSCPIKEDSFLQRY SSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQNPVYHNQPLNPAPSRDPHYQDPHS TAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDNPDYQQDFFPKEAKPNGIFKG STAENAEYLRVAPQSSEFIGA SEQ ID The The nucleotide sequence encoding the heavy NO: 148 nucleotide chain variable region (SEQ ID NO: 36): sequence GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA encoding GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG fusion protein 1 TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG heavy TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 68): ATCCCACCGCACGTTCAGAAGTCGGTTAATAACGACATGATAGTCACTGACAA CAACGGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCACC TGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAG CCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGA GACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCT TCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGT TCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCA GCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO149 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 2 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 69): ATCCCACCGCACGTTAACAACGGTGCAGTCAAGTTTCCACAACTGTGTAAATT TTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAG CATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAA TGACGAGAACATAACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGA CTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCC TGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATC TTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 150 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 3 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 70): ATCCCACCGCACGTTGGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGA TGTGAGATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCAC CTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGA GAACATAACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATT CTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGA GACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAG AAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 151 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 4 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 71): ATCCCACCGCACGTTGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAG ATTTTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATC TGTGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACAT AACACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAA GATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTC TTCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAAT ATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 152 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 5 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 72): ATCCCACCGCACGTTCAACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGT GACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCA CAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGAC AGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCT CCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCC TGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCA ATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 153 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 6 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 73): ATCCCACCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGAC AACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAG GAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTT TGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAA AGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTA GCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCC TGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 154 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 7 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 74): ATCCCACCGCACGTTTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 155 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 8 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 75): ATCCCACCGCACGTTCAGCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGT GACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCA CAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGAC AGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCT CCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCC TGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCA ATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO nucleotide chain variable region (SEQ ID NO: 36): 156 sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 76): ATCCCACCGCACCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 157 nucleotide chain variable region (SEQ ID NO: 36): sequence AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 10 heavy chain TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 77): ATCCCACCGCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT AAA SEQ ID The The nucleotide sequence encoding the heavy NO nucleotide chain variable region (SEQ ID NO: 36): 158 sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC fusion GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA protein 11 GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 78): ATCCCACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO nucleotide chain variable region (SEQ ID NO: 36): 159 sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 12 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 79): ATCCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 160 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 13 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 80): CCACCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAA CCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGA AGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTG CCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAG TGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGC TCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTG ACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 161 nucleotide chain variable region (SEQ ID NO: 36): sequence GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA encoding GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG fusion TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG protein TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC 14 AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC heavy TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC chain ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 81): CCGCACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCA GAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGT CTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCA TGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGC ATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCT GATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACT AAA SEQ ID The The nucleotide sequence encoding the heavy NO: 162 nucleotide chain variable region (SEQ ID NO: 36): sequence GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA encoding GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG fusion TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG protein TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC 15 heavy AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 82): CACGTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAA ATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTG TGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGA CCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATT ATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGAT GAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 163 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion protein GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG 16 heavy TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG chain AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 83): GTTCTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATC CTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGT GGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCC CAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATG AAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAG TGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 164 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACAGTGA fusion AGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG protein TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG 17 heavy AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC chain TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 84): GGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCAC CTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAA GCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAG AGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGC TTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTG TTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACC AGCAATCCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 165 nucleotide chain variable region (SEQ ID NO: 36): sequence GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA encoding GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG fusion TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG protein TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC 18 heavy AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC chain TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 85): GTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGA CAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACA GGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAG TTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCC AAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTG TAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAAT CCTGACTAAA SEQ ID The The nucleotide sequence encoding the heavy NO: 166 nucleotide chain variable region (SEQ ID NO: 36): sequence TGAACACCAAATACAATGAGAAGTTCAAGGGCAGGGTGACCCTGACAGCAGACACC encoding GAGGTCCAACTTGTCCAGTCTGGAGCAGAGGTGAAGAAGCCTGGAGCCACA fusion GTGAAGATTTCCTGTAAGGTGTCTGGCTACACCTTCACCACCTACTACACCCACTGGG protein TGAAGCAGGCTCCTGGCAAGGGATTGGAGTGGATTGGCTGGATTTACCCTGGAGATG 19 heavy AGCACAGACACAGCCTATATGGAACTGTCCTCCCTGAGGTCTGAGGACACAGCAGTC chain TACTACTGTGCCAGGGAGGACCCTGGCAGCAACTACTTTGACTACTGGGGACAAGGC ACCCTGGTGACAGTGTCCAGC The nucleotide sequence encoding the heavy chain constant region (SEQ ID NO: 67): GCAAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG CAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCC AAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACgTGCGTGGTGGTGGACGTG AGCCACGAAGACCCcGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCA TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCG TGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT The nucleotide sequence encoding Linker linking the heavy chain C-terminal and TGFβR2 extracellular domain in the fusion protein (SEQ ID NO: 87): GGTGGTGGCGGTTCAGGCGGAGGTGGCTCTGGAGGTGGAGGTTCAGGAGGT GGTGGTTCT The nucleotide sequence encoding the full- length extracellular domain of TGFβR2 (SEQ ID NO: 86): CTGTGTAAATTTTGTGATGTGAGATTTTCCACCTGTGACAACCAGAAATCCTG CATGAGCAACTGCAGCATCACCTCCATCTGTGAGAAGCCACAGGAAGTCTGTGTGGC TGTATGGAGAAAGAATGACGAGAACATAACACTAGAGACAGTTTGCCATGACCCCAA GCTCCCCTACCATGACTTTATTCTGGAAGATGCTGCTTCTCCAAAGTGCATTATGAAG GAAAAAAAAAAGCCTGGTGAGACTTTCTTCATGTGTTCCTGTAGCTCTGATGAGTGC AATGACAACATCATCTTCTCAGAAGAATATAACACCAGCAATCCTGACTAAA

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1. A truncated TGFβR2 extracellular domain molecule that, compared to its natural form, a) at least the amino acid residues at positions 6-16 thereof are deleted, and further optionally, the amino acid residues at positions 17-17+n thereof are deleted, where n is an integer of 1-10; preferably, n is 2, 4, 8, 9 or 10; most preferably, n is 9; or b) on the basis of the deletion of amino acid residues thereof at positions 6-26, furthermore, the amino acid residues thereof at positions 5, 4-5, 3-5, 2-5, 1, 1-2, 1-3, or 1-4 are deleted; or c) the amino acid residues at positions 7-26 are deleted.
 2. The molecule of claim 1, wherein the amino acid sequence comprises any of SEQ ID NO: 48-62.
 3. A fusion protein comprising the molecule of claim
 1. 4. A fusion protein, comprising a) the truncated TGFβR2 extracellular domain molecule of claim 1; and b) a targeting portion.
 5. The fusion protein of claim 4, wherein the targeting portion is a cancer cell-specific targeting portion selected from an antibody or antigen-binding fragment thereof, a functional ligand or Fc fusion protein thereof, and a receptor protein or Fc fusion protein thereof.
 6. The fusion protein of claim 5, wherein the targeting portion is an anti-EGFR antibody or an antigen-binding fragment thereof.
 7. The fusion protein of claim 4, wherein the N-terminal of the truncated TGFβR2 extracellular domain molecule is linked to the C-terminal of the heavy chain of the targeting portion; and optionally, linked by a linker.
 8. The fusion protein of claim 7, wherein the linker is a G4S flexible peptide linker, preferably a (G4S)4 peptide linker.
 9. An isolated antibody binding to EGFR or an antigen-binding fragment thereof, comprising (a) a heavy chain variable region comprising heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3 domains comprising SEQ ID NOs: 19, 20, and 21, respectively, and/or (b) a light chain variable region comprising a light chain CDR1, light chain CDR2, and light chain CDR3 domain comprising SEQ ID NOs: 16, 17, and 18, respectively.
 10. The antibody or an antigen-binding fragment of claim 9, comprising a) a heavy chain variable region comprising a sequence comprising SEQ ID NO:28 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and/or (b) a light chain variable region comprising a sequence comprising SEQ ID NO: 29 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith.
 11. The antibody or antigen-binding fragment of claim 9, wherein said antibody further comprises: a) a heavy chain constant region, preferably comprising a sequence comprising SEQ ID NO: 30 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and/or b) a light chain constant region, preferably comprising a sequence comprising SEQ ID NO: 31 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith.
 12. The fusion protein of claim 4, wherein the targeting portion is selected from an anti-EGFR antibody of: (a) a heavy chain variable region comprising heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3 domains comprising SEQ ID NOs: 19, 20, and 21, respectively, and/or (b) a light chain variable region comprising a light chain CDR1, light chain CDR2, and light chain CDR3 domain comprising SEQ ID NOs: 16, 17, and 18, respectively, Trastuzumab, Bevacizumab, Ramucirumab, Ipilimumab, or Panitumumab.
 13. The fusion protein of claim 7, wherein a) the amino acid sequence of a heavy chain comprises SEQ ID NO:141 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; and b) the amino acid sequence of a light chain comprising SEQ ID NO:23 or a sequence having at least 85%, 88%, 90%, 95%, 98%, or 99% sequence identity therewith; wherein the fusion protein comprises two heavy chains and two light chains; a disulfide bond is formed between a first light chain and a first heavy chain thereof, a disulfide bond is formed between a second light chain and a second heavy chain thereof, and a disulfide bond is formed between a first heavy chain and a second heavy chain thereof.
 14. The fusion protein of claim 13, having a KD value of 2.92 pM-26.3 pM, preferably 7 pM-9 pM, most preferably 8.77 pM for binding affinity to human EGFR protein, and having a KD value of 23 pM-288.3 pM, preferably 64 pM-144 pM, most preferably 96.1 pM for binding affinity to human TGF-β1 protein.
 15. A conjugate comprising the truncated TGFβR2 extracellular domain molecule of claim 1 and an additional therapeutic agent, preferably said antibody or antigen-binding fragment thereof and the additional therapeutic agent are linked by a linker.
 16. A nucleic acid encoding the truncated TGFβR2 extracellular domain molecule of claim 1, which is mRNA and/or DNA.
 17. The nucleic acid of claim 16, comprising any one of SEQ ID NOs: 32 to 39; or any one of SEQ ID NOs: 67-84; or any one of SEQ ID NOs: 148-163, or an functional variant thereof.
 18. An expression vector comprising the nucleic acid of claim
 16. 19. A host cell comprising the nucleic acid of claim
 16. 20. A method for producing the truncated TGFβR2 extracellular domain molecule of claim, comprising culturing a host cell comprising a nucleic acid encoding the truncated TGFβR2 extracellular domain molecule of claim 1, which is mRNA and/or DNA, under conditions suitable for the expression of the preceding protein molecule, and recovering the expressed product from the culture medium.
 21. A pharmaceutical composition comprising a) the truncated TGFβR2 extracellular domain molecule of claim 1, and b) a pharmaceutically acceptable carrier; optionally c) one or more additional therapeutic agents.
 22. Use of the truncated TGFβR2 extracellular domain molecule of claim 1 for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.
 23. Use of the truncated TGFβR2 extracellular domain molecule of claim 1 for the preparation of a medicament for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.
 24. A pharmaceutical combination comprising the truncated TGFβR2 extracellular domain molecule of claim 1 and one or more additional therapeutic agent(s).
 25. A kit comprising the truncated TGFβR2 extracellular domain molecule of claim 1, further comprising a device for administering the medicament.
 26. A method of preventing and treating a neoplastic disease comprising administering to a subject the truncated TGFβR2 extracellular domain molecule of claim
 1. 27. A pharmaceutical composition comprising a) the fusion protein of any one of claim 3, and b) a pharmaceutically acceptable carrier; optionally c) one or more additional therapeutic agents.
 28. A pharmaceutical composition comprising a) the nucleic acid of claim 16, and b) a pharmaceutically acceptable carrier; optionally c) one or more additional therapeutic agents.
 29. Use of the fusion protein of claim 3 for the prevention and treatment of cancer or for the preparation of a medicament for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.
 30. Use of the nucleic acid of claim 16 for the prevention and treatment of cancer or for the preparation of a medicament for the prevention and treatment of cancer, preferably for the treatment of gastric cancer.
 31. A pharmaceutical combination comprising the fusion protein of claim 3, and one or more additional therapeutic agent(s).
 32. A pharmaceutical combination comprising the nucleic acid of claim 16, and one or more additional therapeutic agent(s).
 33. A pharmaceutical combination comprising the pharmaceutical composition of claim 21, and one or more additional therapeutic agent(s).
 34. A kit comprising the fusion protein of claim 3; further comprising a device for administering the medicament.
 35. A kit comprising the nucleic acid of claim 16; further comprising a device for administering the medicament.
 36. A kit comprising the pharmaceutical composition of claim 21; further comprising a device for administering the medicament.
 37. A method of preventing and treating a neoplastic disease comprising administering to a subject the fusion protein of claim
 3. 38. A method of preventing and treating a neoplastic disease comprising administering to a subject the nucleic acid of claim
 16. 39. A method of preventing and treating a neoplastic disease comprising administering to a subject the pharmaceutical composition of claim
 21. 